MXPA06005950A - Biomarkers for the efficacy of calcitonin and parathyroid hormone treatment. - Google Patents

Biomarkers for the efficacy of calcitonin and parathyroid hormone treatment.

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MXPA06005950A
MXPA06005950A MXPA06005950A MXPA06005950A MXPA06005950A MX PA06005950 A MXPA06005950 A MX PA06005950A MX PA06005950 A MXPA06005950 A MX PA06005950A MX PA06005950 A MXPA06005950 A MX PA06005950A MX PA06005950 A MXPA06005950 A MX PA06005950A
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gene expression
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MXPA06005950A
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Maria Bobadilla
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Novartis Ag
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Abstract

A mufti-organ gene profiling analysis of the results of an administration to a subject of salmon calcitonin or a parathyroid hormone analogue provides biomarkers of calcitonin treatment efficacy and parathyroid hormone or parathyroid hormone analogue treatment efficacy. Among the biomarkers are the expression profiles of the genes for Y-box binding protein, BMPs, FGFs, IGFs, VEGF, alpha-2-HS glycoprotein (AHSG), OSF, nuclear receptors (steroid/thyroid family) and others. The results obtained support the anabolic effect of salmon calcitonin on bone metabolism.

Description

BIOM ARCADO RES TO DETERMINE THE EFFECTIVENESS OF THE TREATMENT WITH CALCITONIN AND HORMONE PARATIROIDES FIELD OF THE INVENTION This invention relates in general to the analytical testing of tissue samples in vitro, and more particularly to aspects of the profiling of gene expression with respect to calcium regulation.
BACKGROUND OF THE INVENTION Calcium is essential for many cellular processes in the body, and is especially important for bone metabolism. The level of calcium in the body is carefully maintained by an endocrine control system. Two of the hormones in this endocrine control system are calcitonin and the parathyroid hormone. Calcitonins, which are polypeptide hormones of approximately 32 amino acids, are an endogenous regulator of calcium homeostasis, and can be used as anti-resorption agents for the treatment of disorders associated with hypocalcemia. Calcitonin is produced in parafollicular cells (C cells) of the thyroid gland. Several calcitonins, including, for example, salmon and eel calcitonin, are commercially available, and are commonly used in the treatment, for example, of Paget's bone disease, malignant hypocalcemia, and post-menopausal osteoporosis. Pondel Intl. J. Exp. Pat ol. 81 (6): 405-22 (2000). A version of calcitonin (Miacalcin®) is available as a nasal spray. Parathyroid hormone (PTH) is a polypeptide of 84 amino acids. Parathyroid hormone regulates bone remodeling and Ca2 + homeostasis. Parathyroid hormone is also a paracrine activator known for the differentiation and activity of osteoclasts. PTS893 [SDZ PTS 893; Leu8, Asp10, Lys11, Ala16, Gln18, Thr33, Ala34, human PTH 1-34 [h PTH (1-34)]], is a 34 amino acid parathyroid analog that improves bone mass and biomechanical properties. Kneissel M. and collaborators, Bone 28: 237-50 (March 2001); Stewart A. F. et al., J. Bone. Miner Res. 15 (8): 1517-25 (August 2000), Thomsen J. S. et al., Bone 25 (5): 561-9 (November 1999). It is known that calcitonin and parathyroid hormone interact in a complex and interdependent manner, but understanding the way in which calcitonin and parathyroid hormone interact has been incomplete. The inhibitory effects of calcitonin on the resorption activity of the osteoclasts and on the renal tubular calcium resorption have been well documented.
However, the potential effects of calcitonin on osteoblasts and interactions with any other factors related to skeletal metabolism have remained controversial. An analysis of genetic profiling of multiple organs would provide a better picture of the changes induced by a compound over the whole organism, and would also give a new perspective to the understanding of the pharmacology of the hormones. Genomic technologies are a source of the new hypothesis-generating capabilities that are now empowering biomedical researchers. In the context of drug development, they provide a new perspective to understanding the pharmacology of drugs. In accordance with the foregoing, there is a need in the art for an understanding of the whole organism on the activity of calcitonin and parathyroid hormone.
COMPENDIUM OF THE INVENTION The invention provides an answer to the need of the technique. The multi-organism gene profiling analysis provides a complete picture of the changes induced by a compound throughout the organism, and gives a new perspective to the understanding of drug pharmacology. In one aspect, the invention provides the first description of the molecular mechanisms of action of hormone-mediated bone remodeling by salmon caletonin, by means of genetic profiling analysis. The known mechanisms of action of caletonin as an anti-resorption agent could be reconstructed at the molecular level. Effects on effectors and pathways linked to bone remodeling activities - BMPs were also observed, IGFs, matrix components ext ra ce I u I a r, and VEGF -. These results support the role of calcitonin as an anabolic agent. In another aspect, the invention provides the first reconstruction of the molecular mechanisms of action of a pharmacological agent on one of its objective tissues in an intact primate animal model, by evaluating changes in gene expression induced by salmon calcitonin or by the analogue of parathyroid hormone PTS893 on the bones of cynomolgus monkeys, in order to elucidate the molecular mechanisms of action that mediate their effects. The analysis of genetic profiling allowed the reconstruction of the pathways involved in the transduction of the calcitonin signal, triggered by the stimulation of the G-protein-linked receptor, and its influence on the cell cycle, as indicated by the changes observed in the cyclins. In vivo genetic expression expression studies allow the identification of the molecular mechanisms underlying a pharmacological effect. In one embodiment, the invention provides the use of calcitonin in the manufacture of a medicament for the treatment of a condition for which an anabolic agent is indicated. In one embodiment, the condition is atherosclerosis. The invention also provides the use of calcitonin in the manufacture of a medicament for the treatment of calcium metabolism disorders in a selected patient population, wherein the patient population is selected based on the gene expression profile indicating the efficacy of calcitonin by the patient to whom the calcitonin is administered. In one embodiment, calcitonin is salmon calcitonin. The invention further provides the use of a parathyroid hormone or a parathyroid hormone analogue in the manufacture of a medicament for the treatment of calcium metabolism disorders in a selected patient population, wherein the patient population is selected based on the genetic expression profile that indicates the effectiveness of the parathyroid hormone or the parathyroid hormone analogue by the patient receiving the parathyroid hormone or the parathyroid hormone analogue. In one embodiment, the parathyroid hormone analogue is PTS893. In one embodiment, the drug is administered in a therapeutic dose before determining the gene expression profile by the patient. In another embodiment, the medicament is administered in a sub-therapeutic dose before determining the gene expression profile by the patient. The invention also provides a method for treating a condition in a subject, wherein the condition is one for which the administration of a calcitonin, parathyroid hormone, a parathyroid hormone analog, or a combination thereof is indicated. The method involves first administering a compound of interest to the subject (e.g., a primate subject), and then obtaining the gene expression profile of the subject following administration of the compound. The gene expression profile of the subject is compared to a gene expression profile of a biomarker. The gene expression profile of the biomarker indicates the efficacy of the treatment with a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination thereof. In one embodiment, the gene expression profile of the biomarker is the gene expression profile of the baseline of the subject prior to administration of the compound. In another modality, the genetic expression profile of the biomarker is the gene expression profile or the average of the gene expression profiles of a vertebrate that has been administered cale i ton i na (for example, salmon calcitonin), or hormone parathyroid, or a parathyroid hormone analog (for example, PTS893). A similarity in the gene expression profile of the subject to which the compound was administered, with the gene expression profile of the biomarker, indicates the efficacy of the treatment with the compound. In accordance with the foregoing, the invention provides biomarkers for determining the efficacy of the treatment of a condition for which calcitonin, parathyroid hormone, or a combination thereof is indicated. Biomarkers include the expression profiles of the genes for the Y-frame binding protein, bone morphogenetic proteins (BMPs), fibroblast growth factors (FGFs), insulin-like growth factors (IGFs), vascular endothelial growth factor (VEGF), glycoprotein a-2-HS (AHSG), osteoclast stimulating factor (OSF), nuclear receptors (steroid family is / ti ro ides), and others. The invention provides methods for determining a subject to be included in a clinical study, based on an analysis of the biomarkers expressed in the subject to be treated. The subject to be tested is administered to the subject. In one embodiment, the compound to be tested is administered in a sub-therapeutic dose. Then the gene expression profile of the subject is obtained following the administration of the compound. The subject can be included in the clinical study when the gene expression profile of the subject to which the compound was administered is similar to a gene expression profile of a biomarker that indicates the efficacy of treatment with a calcitonin, parathyroid hormone, an analog of parathyroid hormone, or a combination thereof. The subject can be excluded from the clinical study when the genetic expression profile of the subject is different from the biomarker gene expression profile that indicates the efficacy of the treatment. These similarities or differences can be observed by experts in this field. The invention also provides clinical trials, kits of parts, and reagents to determine the effectiveness of the treatment of a condition for which the administration of a calcitonin, a parathyroid hormone, or a parathyroid hormone analogue is indicated. In one embodiment, the kits of parts contain reagents for determining the genetic expression of the biomarker genes, by hybridization. In another embodiment, the kits of parts contain reagents to determine the genetic expression of the biomarker genes, by means of the chain reaction of the polymerase.
DESCRIPTION OF THE PREFERRED MODALITIES This invention is based on the understanding of the effects of administration to a subject of calcitonin (eg, salmon calcitonin, SEQ ID NO: 1) or parathyroid hormone (SEQ ID NO: 2), or a analogue thereof (eg, PTS893; SEQ ID NO: 3). An analysis of multi-organ genetic profiling of the results of administration to a subject of salmon calcitonin or of a parathyroid hormone analogue provides biomarkers of the efficacy of treatment with calcitonin and of the effectiveness of treatment with parathyroid hormone or with a parathyroid hormone analog. As used herein, a subject is a vertebrate. In one modality, the vertebrate is a mammal. In a more particular embodiment, the subject is a primate, for example a cynomolgus monkey or a human being. The present analysis, describes globally the molecular mechanisms of action of salmon calcitonin and PTS893 to change the content of ribonucleic acid (RNA) in different organs, through the genetic profiling analysis of multiple organs in primates. The RNA content of the cell, the "transcriptomo" is a reflection of the functions and the state of the cell. Within an individual cell or an organ, the expressions of the different elements of a transcriptomo are not independent. The change in the level of expression can trigger a series of events that will eventually lead to another modification of the transcriptomo. These interdependent events are described in terms of paths. Because changes in the different functions within a cell are closely interconnected, changes in different organs within the organism are linked. The application of genetic profiling to different organs subjected to the same treatment gives a better overview of the effects and changes in the physiological state. As shown herein, this is particularly the situation when the profiling analysis of multiple organs of the compounds p I and i or t r or p i c s, such as calcitonin, is to be carried out. In fact, the overall signature described for calcitonin is reflected not only in the main target organ (ie bone), but also in the other organs analyzed in the present. In this multi-organ genetic profiling analysis, the known mechanisms of action of calcitonin as an anti-resorptive agent and of the parathyroid hormone PTS893 as a paracrine activator of the differentiation and activity of osteoclasts, could be reconstructed at the level molecular. The inhibitory effect of calcitonin on osteocytes could be reconstructed, with changes that affect, among others, the genes for PU.1 (SPI1; SpiB; SEQ ID NO: 4), the colony stimulating factor (CSF-1). (SEQ ID NO: 6), to differentiation and survival); to carbonic anhydrase (SEQ ID NO: 8), H + -ATPases, cathepsin K (resorption activity), tubulins, PAK4 (mobility). The effects on effectors and the pathways linked to bone remodeling activities (bone morphogenetic proteins (B Ps), fibroblast growth factors (FGFs), insulin-like growth factors (IGFs), extracellular matrix components, were also observed. spheroid hormones, vascular endothelial growth factor (VEGF), and glycoprotein a-2-HS (AHSG), shared in many cases by both salmon calcitonin and PTS893.It is interesting that salmon calcitonin It also regulates the expression of genetic coding for osteocyst stimulating factor (OSF), and cystatin.It is also interesting that PTS893 also regulates the genes involved in the differentiation and survival of osteocytes (SPM, CSF-1, protein associated with the differentiation of monocytes to macrophages (MD).) PTS893 also produced a sharp increase in nuclear receptors (steroid / thyroid family). With the above, these results support the role of calcitonin as an anabolic agent.
Calcitonin is currently used in the treatment of systemic skeletal diseases characterized by high bone mass, which are a consequence of the imbalance between bone formation (anabolic) and bone resorption, the former predominating. Calcitonin promotes the synthesis of bone morphogenetic protein-2 (BMP-2), which is known to be a potent anabolic agent. There is strong evidence that, when calcitonin reaches bone cells, these can have an anabolic effect by increasing BMP-2 production. Accordingly, calcitonin can be used in a method for the treatment of an individual in order to adjust the bone mineral density of a subject. This is the first approach to characterize, in an in vivo model, the effects of calcitonin on bone metabolism, by profiling gene expression. The inhibitory effect of calcitonin on osteoclasts could be reconstructed, with changes that affect the genes such as carbonic anhydrase, H + -ATPases, and cathepsin K. Salmon calcitonin also appeared to regulate the expression of the gene that codes for cystatin, being this effect described in the present for the first time. Salmon calcitonin also has modulating effects on the genes that affect the direct, autocrine, paracrine, and endocrine regulation of mesenchymal cell functions, such as pleiotropin, periostin, fibro-growth factor, transforming growth factors-betas (TGF-betas), insulin-like growth factor / binding proteins (IG Fs / IGFB Ps), bone morphogenetic proteins (B Ps), Vascular Endothelial Growth Factor (VEGF), Tumor Necrosis Factor (TNF), neurocondrine, type 3 folistatin, or parathyroid hormone receptor. It also regulates the synthesis and degradation of extracellular matrix components (collagens, osteopontin, osteocalcin, dermatopontine, chondroadherin, glypican, or syndecan), and enzymes. Salmon calcitonin also influences some aspects of bone mineralization, due to changes in dentin. As provided herein, calcitonin can also be used as an anabolic agent in the treatment of other conditions where anabolism or tissue growth is therapeutically desirable. This condition is atherosclerosis, an atheromatous disease where atheromatous plaque is complicated by fibrosis and calcification. Moreover, the invention provides biomarkers of the efficacy of treatment with calcitonin or parathyroid hormone. As used herein, a gene expression profile is a diagnosis to determine the effectiveness of the treatment when the gene expression increased or decreased is an increase or decrease (eg, at least a difference of 1.5 times) on gene expression of the baseline following the administration of the compound (ie, the gene expression profile of the biomarker is the gene expression profile of the baseline of the subject prior to administration of the compound). Alternatively or in addition, the gene expression profile is the diagnosis to determine the efficacy of the treatment, compared to the treatment of calcitonin (eg, salmon calcitonin), or parathyroid hormone, or parathyroid hormone analogues ( for example, PTS893), when the gene expression profile of the treated subject can be compared with a gene expression profile of a standard biomarker. In one embodiment, the gene expression profile of the standard biomarker is the gene expression profile or the average gene expression profiles of a vertebrate that has been administered a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination of them, this profile being the standard with which the subject's results are compared with the administration. This approach, which contains aspects of therapy and diagnosis, is termed as "teranostic" by many experts in this field.
In one modality, the subject is a vertebrate. In a particular embodiment, the vertebrate is a mammal. In a more particular embodiment, the mammal is a primate, such as a cynomolgus monkey or a human being. As used herein, the administration of an agent or a drug to a subject or patient, includes self-administration or administration by another. As used herein, a gene expression profile is the diagnosis of the efficacy of the 10 treatment with calcitonin or with parathyroid hormone when the increased or decreased gene expression is an increase or decrease (for example, at least a difference of 1.5 times) on the genetic expression of the baseline following the administration of a 15 calcitonin, or a parathyroid hormone or an analog. As used herein, a pattern of gene expression is "higher than normal" when genetic expression (eg, in a sample of a treated subject) shows a difference of 1.5 times (ie, higher) in 2o the level of expression, comparing with the samples of the baseline. A gene expression pattern is "lower than normal" when genetic expression (for example, in a sample of a treated subject) shows a difference of 1.5 times (ie, lower) in the level of expression, "Comparing with the baseline samples.
Techniques for the detection of gene expression of the genes described by this invention include, but are not limited to, Northern blots, RT-PCT, real-time polymerase chain reaction, primer extension, RNAse protection, profiling of RNA expression, and related techniques. Techniques for detecting gene expression by detecting the protein products encoded by the genes described by this invention include, but are not limited to, antibodies that recognize protein products, Western blots, immunofluorescence, immunoprecipitation, ELiSAs, and related techniques. These techniques are well known to those skilled in the art. Sambrook J. et al., Molecular Cloning: A Laboratory Manual, Third Edition, (Cold Spring Harbor Press, Cold Spring Harbor, 2000). In one embodiment, the technique for detecting gene expression includes the use of a genetic chip. The construction and use of the genetic chips are well known in the art. See Patents of the United States of North America Nos. 5,202,231; 5,445,934; 5,525,464; 5,695,940; 5,744,305; 5,795,716, and 5,800,992. See also, Johnston, M. Curr. Biol. 8: R171-174 (1998); lyer V. R. et al., Science 283: 83-87 (1999) and Elias P. "New human genome 'chip' is a revolution in the offing" Los Angeles Daily News (October 3, 2003).
The gene expression profile may include one or more genes selected from the isoform a group of acid phosphatase 1; 1 receptor type of activin A, type II; Activin A receptor type IIB precursor; activin beta-C chain; alpha 2 HS glycoprotein; amelogenin; annexin V; arylsulfatase precursor E; ATPase H (+) vacuolar; ATPase H (+), vacuolar subunit; ATPase, H + transport, lysosomal; ATPase, transport of H +, lysosomal; ATPase, transport of H +, lysosomal; bigücano; bone morphogenetic protein 1; orfogenic bone protein 10; bone morphogenetic protein 2A; bone morphogenetic protein 5; precursor of bone morphogenetic protein 6; Calcium binding protein 1 (calbrain); calcium-dependent protein / calmodulin kinase (CaM kinase) II gamma; calreticulin; element modulator that responds to cAMP (CREM); carbonic anhydrase I; carbonic anhydrase II; precursor of oligomeric cartilage matrix protein; cathepsin K; cathepsin W; kinase 1 type CDC; kinase 2 type CDC, isoform hclk2 / 139; chondroitin sulfate proteoglycan 2 (versican); proteoglycan 3 of chondroitin sulfate (neurocan); chorionic somatomammotropin hormone 1; Chymotrypsin C (Caldecrine); type 1 collagen and fusion transcription of PDGFB; collagen type II alpha 1; collagen type III alpha 1; collagen type IV alpha 2; type IX alpha 1 collagen; collagen type VI alpha 1; collagen type VI alpha 2 (AA 570 998); collagen type XI alpha 1; collagen type XI alpha 2; collagen type XI alpha 2; collagen, type I, alpha 2; collagen type IV, alpha 1; collagen type IX, alpha 2; collagen type V, alpha 2; collagen type VI, alpha 1; precursor of collagen type VI, alpha 1; collagen type XVI, alpha 1; collagen type XVI, alpha 1; collagenase 3 (matrix metalloproteinase 13); connective tissue growth factor; cyclin A2; Cyclin B1; Cyclin D2; cyclin E2; cyclin-dependent qulphan 5; cyclin-dependent kinase 5, regulatory subunit 1 (p35); cyclin-dependent kinase 6; cyclin 1A-dependent kinase inhibitor (p 21, C i p 1); Cystatin B (Stefin B); cytokine-inducible kinase; protein kinase 1 associated with death; protein kinase 3 associated with death; f osf or roteí n a acid of dentin matrix 1 (DMP1); double specificity phosphatase 9; myotonic protein kinase of dystrophy; pyrophosphatase / ecto-nucleotide phosphodiesterase 1; pyrophosphatase / ecto-nucleotide phosphodiesterase 1; endoteiial differentiation, precursor of receptor coupled with G6 protein, estrogen receptor; estrogen receptor; protein related to estrogen receptor; B-box protein that responds to estrogen (EBBP); fibroblast activation protein; fibroblast growth factor 1 (acid); fibroblast growth factor 18; fibroblast growth factor 4; fibroblast growth factor receptor; Folistatin type 1; Folistatin type 1; glutamate receptor, metabotropic 1; component. of acetyl-glucosaminyl transferase GPI1 N, g p 1 1; granulocyte macrophage colony stimulating factor (CSF1); inducible by growth arrest and DNA damage, alpha; protein 10 linked to growth factor receptor; proteoglycan of heparan sulfate 2 (perlecan); inositol 1, 4, 5-triphosphate receptor, type 1; inositol 1, 4, 5-triphosphate receptor, type 1; inositol 1, 4, 5-triphosphate receptor, type 2; kinase 3 isoenzyme of inositol 1,4,5,5-triphosphate; inositol 4 type I beta polyphosphate phosphatase; Inositol 5 polyphosphate phosphatase; inositol monophosphatase 1 (myo) 1 (or 4), 1; inositol monophosphatase 2 (my) 1 (or 4); insulin-like growth factor (IGF II); insulin-like growth factor 2 (somatomedin A); insulin-like growth factor binding protein; Insulin-like growth factor-2 protein; insulin-like growth factor protein 3; enl lace protein 5 of insulin-like growth factor; insulin protein growth factor 2 protein; insulin-like growth factor II precursor; insulin-like growth factor II precursor; alpha-10 subunit of integrin; kinase associated with interleukin 1 receptor; Janus 3 kinase; LIM protein (similar to the binding enigma of rat protein kinase C); Lysyl oxidase type protein; MAD, homologue 3 of mothers against decapentaplégico; MAGUKs (membrane associated guanylate kinase homologs; AP kinase kinase kinase (MTK 1); MAPK13; mitogen-activated protein kinase 13; MAPK8IP1: protein 1 interaction with mitogen-activated protein 8 kinase; EK kinase; metalloproteinase, mitogen-activated protein kinase 1, mitogen-activated protein kinase 8, mitogen-activated protein kinase kinase 1, mitogen-activated protein kinase kinase kinase 4 kinase, protein kinase-activated kinase 2 mitogen-activated protein; protein kinase 3 activated by mitogen-activated protein kinase; MDD: associated with monocyte to macrophage differentiation; neurocondrine; nuclear factor of activated T cells, cytoplasmic, dependent on calcineurin 1; OS 4 protein (OS 4); osteoblast-specific factor 2 OSF 2 (periostin); osteoclast stimulating factor (OSF); PAK4; protein associated with PDFG; Phosphatidylinositol 4 kinase, catalytic, beta polypeptide; f or sf at i d i I n o n t i I g g I n g, class L; Phosphatidylinositol polyphosphate phosphatase 5, isoform B; Phosphatidylinositol 4 phosphate kinase 5, C (1) isoform; Phosphatidylinositol 4 phosphate kinase 5, type I, beta; Phosphatidylinositol 4 phosphate kinase 5, type II, beta; phosphatidylinositol-glycan, class C (PIG C); f osfodiesterase 4A, specific for cAMP; phosphodiesterase 4D, specific for cAMP (f osph od iesterase E3 homologous dunce { Drosophila)); Phosphodiesterase IB, calmodulin-dependent; phosphoinositide 3 kinase of phosphoinositide 3, catalytic, phosphoinositide 3, class 3, gamma-kinase polypeptide; phospholipase C b3 phospholipase C, beta 4; phospholipase D; protein transfer of f or sf ati d i I n t i I; protein D2 kinase PKD2; pre-pro-collagen type!, alpha 2; pre-pro-collagen type I, alpha 1; pro-collagen alpha 1, type II; Procollagen Lysine Dioxygenase 5; procollagen-proline, 2-oxoglutarate-4-dioxygenase (proline hydroxylase 4), alpha I polypeptide; endometrial protein associated with progestogen (placental protein 14, endometrial alpha 2 globulin associated with pregnancy, 'alpha uterine protein'); prolidase (imido-dipeptidase) PEPD; proliferating cell nuclear antigen; prolyl hydroxylase 4 beta; protease, serine, 11 (IGF binding); Proteasome subunit (prosoma, macropain), beta type, 10; activated STAT X protein inhibitor; protein kinase 1 PCTAIRE; 80K H substrate protein kinase C; protein kinase C, alpha; protein kinase, cAMP dependent, catalytic, gamma; protein kinase, cAMP-dependent, regulatory, type I, beta; protein kinase, dependent on cAMP, regulatory, type II, alpha; P2Y purinergic receptor, coupled with G protein, 11; substrate 2 of botulinum toxin C3 related to RAC2 Ras (rho family, small GTP binding protein Rac2); receptor tyrosine kinase DDR; retinoid receptor X, gamma; protein kinase kidney osome S6; ribosomal protein kinase S6, 9kD, polypeptide 3; SCAMP1: secretory carrier membrane protein 1 (vesicular transport); f osphoprotein secreted 1 (osteopontin, if a I or p rote i n to bone I, activation of early T-lymphocytes 1); serine (or cysteine) proteinase inhibitor, cluster H (heat shock protein 47), member 2; serine / threonine kinase 38; serine / threonine protein kinase; SF 1; steroidogenic factor 1; signal transducer and transcription activator 1; signal transducer and transcription activator 2, 113 k D; signal transducer and transcription activator 5A; signal transducer and transcription activator 5A; signal transducer and transcription activator 6 (STAT6); Smad 3; Smad anchor for receptor activation, soforma 1; Smad5; SMAD6 (inhibits B P / Smad1 (MADH1)); kinase related to SNF1; transcription factor SpiB (related to SPI1 / PU.1); Stat5b (stat5b); serine / threonine kinase related to Ste20 TEIG; early growth response inducible by TGFB early growth response inducible by TGFB; TIEG anti-apoptotic factor 1 induced by TGFB1; 12 apoptosis protein induced by TGF beta; TGF beta precursor; TGF beta superfamily protein; Tob; Demellated type 1 kinase; transforming growth factor receptor, beta III (betaglycan, 300kD); Transforming growth factor beta 3 (TGF beta 3); TRIO: triple functional domain (interaction with PTPRF); tubulin alfa 1; tubulin alfa 3; tubulin alfa, H2 alpha isotype; tubulin beta 2; tubulin beta 3; tubulin beta 4; tubulin beta, cofactor D; alpha-2 chain precursor of type VI collagen; ubiquitin carrier protein E2 C; Vascular endothelial growth factor; Vascular endothelial growth factor; Vascular endothelial growth factor B; and Y-frame link protein 1. As used herein, the administration of an agent or a drug to a subject or patient, includes self-administration and administration by another. Calcitonin The term "calcitonin" includes not only the naturally occurring caicyonins, but also their pharmaceutically active derivatives and analogs, for example, wherein one or more of the peptide residues present in the naturally occurring product are replaced, or where the term N or C is modified. The preferred caicyonins to be used according to the invention are the salmon caicyonins., human, and porcine, and Elcatonina. All of these compounds are commercially available, and have been described extensively, together with their pharmaceutical properties, in the literature. See Patents of the United States of North America Nos. 5,733,569 and 5,759,565, the content of which is incorporated by reference. The amount of calcitonin to be administered according to the method of the invention, and consequently, the amount of active ingredient in the composition of the invention, depends on the particular calcitonin selected, on the condition to be treated, of the desired administration frequency, and the desired effect. The bioavailability for calcitonins, in particular for salmon calcitonin, as determined in terms of concentration in blood plasma following nasal administration, is high, generally on the order of about 50 percent of the levels reached with the intramuscular injection. Accordingly, the administration according to the invention will be carried out appropriately so as to give a dosing index of the order of two times or more, for example of approximately two to four times, the dosing index required for the treatment by means of of intraparietal administration, for example intramuscular. Information regarding the administration of Micalcín® (salmon calcitonin) nasal spray, is available in the prescription information of M iacalcin® (Novartis, November 2002). For intramuscular injection, individual dosages of about 50 to 100 MRC units are applied, at an index of about once a day to about three times a week. For nasal administration according to the. According to the invention, therefore, the treatment will suitably comprise administering dosages from about 50 to about 400 MRC units, more preferably from about 100 to about 200 MRC units, at a frequency from about once a day to about three times a week. Conveniently, the aforementioned dosages will be administered in a single application, that is, the treatment will comprise the administration of individual nasal dosages comprising from about 50 to about 400 MRC units, preferably from about 100 to about 200 MRC units. calcitonin. Alternatively, these dosages can be divided over a series, for example, from two to four applications taken at intervals during the day, the dosage then comprising in each application from about 10 to about 200 MRC units, preferably about 25 units. to approximately 100 MRC units.
The amount of the total composition administered in each nasal application suitably comprises from about 0.05 to 0.15 milliliters, typically about 0.1 milliliters, for example 0.09 milliliters. The compositions for use in accordance with the foregoing, suitably comprise from about 150 to about 8,000, preferably from about 500 to about 4,000, more preferably from about 500 to about 2,500, and most preferably from about 1,000 to about 2,000 units. MRC calcitonin, for example salmon calcitonin, per milliliter. The term "calcitonin" also encompasses active peptide analogs and mimetics, such as are described, for example, in U.S. Patent Nos. 5,719,122, 5,175,146, and 5,698,6721. See U.S. Patent Application Number 2003015815. The "calcitonin superfamily" consists of calcitonin, a peptide related to the calcitonin gene (CGRP), and amylin. Calcitonin and CGRP are derived from the CT / CGRP gene in humans. An alternative splicing of the transcription of the primary RNA leads to the translation of the CGRP and CT peptides in a tissue-specific manner. CGRP (a neuropeptide of 37 amino acids) and its receptors, are widely distributed in the body. Amylin (a 37 amino acid peptide) is generated from a gene located on chromosome 12 (it is thought to be an evolutionary duplication of chromosome 11) and shares a homology of 46 percent amino acid sequence with CGRP, and of 20 percent with human calcitonin. The term "peptide related to the calcitonin gene" or "CGRP" includes the native CGRP, preferably the human CGRP, and its active analogues. It is known that CGRP has a variety of roles in the formation of bones. The term "amylin" includes native amylin, typically from a human source, and pharmaceutically active analogues thereof. It is known that the hormone induces the formation of bone mass through a variety of mechanisms. "Calcitonin-like agents" include "calcitonin", "CGRP", and "amylin". See U.S. Patent Application Number 003015815. Parathyroid hormone. The term "parathyroid hormone" refers to the parathyroid hormone, to fragments or metabolites thereof, and to structural analogues thereof, which can stimulate bone formation and increase bone mass. Also included are peptides related to the parathyroid hormone, and the active and analogous fragments of the peptides related to the parathyroid hormone. See Patents of the United States of North America Nos. 4,086,196; 5,001,223; 6,541, 450, and 6,649,657, and published PC Patent Applications Nos. WO 94/01460 and WO 93/06845. The functional activity of the parathyroid hormone is easily determined by experts in this field, according to conventional tests. A variety of these compounds are described and referenced below; however, other parathyroid hormones will be known to those skilled in the art. Exemplary parathyroid hormones are disclosed in the references cited in U.S. Patent Nos. 6,541, 450 and 6,649,657, the entire contents of which are incorporated by reference. The utility of parathyroid hormones as medical agents in the treatment of conditions that present with low bone mass (eg, osteoporosis) in mammals, is demonstrated by the activity of parathyroid hormones in conventional assays, including in vivo assays , receptor binding assay, cyclic AMP assays, and fracture healing assays. PTS893 is an analogue of the endogenous parathyroid hormone, where certain sites of chemical instability within the N-terminal parathyroid hormone fragments are eliminated., making substitutions of appropriate amino acids in the particular residues, which results in stable and biologically active human parathyroid hormone fragments. The N-terminal fragments of human parathyroid hormones include the hPTH (1-34) OH muteins and the hPTH (1 -38) OH muteins. PTS893 comprises at least the first 27 units of N-terminal amino acids of the parathyroid hormone. Preferred parathyroid hormone derivatives are those which comprise at least one amino acid unit replaced in one or more of the following positions of the parathyroid hormone sequence: 8-11, 13, 16-19, 21, 22, 29 to 34 , in particular 8-11, 16-19, 33 and / or 34. These compounds exhibit desirable bone-forming properties, both in vivo and in vitro, that are equal to, or above, the level of the natural parathyroid hormone. and its N-terminal fragments. See European Patent Number EP 0,672,057; Patent Application of the published TCP Number WO 94/02510; Kneissel M. and collaborators, Bone 28: 237-50 (March 2001); Stewart A. F. and collaborators, J, Bone Miner. Res. 15 (8): 1517-25 (August 2000); Thomsen J. S. et al., Bone 25 (5): 561-9 (November 1999). Cases of parts. The kits of parts of the invention may contain a product written on or inside the case container. The written product describes how to use the reagents contained in the kit of parts, for example, to determine whether a patient is effeely responding, or can effeely respond, to a compound to be used in the treatment of a condition for which Indicate calcitonin, parathyroid hormone, a parathyroid hormone analog, or a combination thereof. In various embodiments, the use of the reagents may be in accordance with the methods of the invention. In one embodiment, the reagent is a genetic chip to determine the gene expression of the relevant genes. The following Example is presented for the purpose of more fully illustrating the preferred embodiments of the invention. This Example is in no way to be construed to limit the scope of the invention, as defined by the appended claims.
EXAMPLE CALCITONINA DE SALMÓN AND PTS893, PHARMACOGENOMIC EXPLORATORY STUDY IN MONOS; ANALYSIS OF GENETIC EXPRESSION OF MICROARREGLO. Introduction and summary. The purpose of this Example was to evaluate changes in gene expression in cynomolgus monkeys following a two-week subcutaneous treatment with salmon calcitonin (sCT) at 50 micrograms / animal / day, and PTS893 at 5 micrograms / animal day, with the In order to elucidate the mechanisms of action that mediate their effects, as well as for the identification of the biomarkers of therapeutic indications. It is believed that this Example is the first analysis that globally describes the molecular mechanisms of action of salmon calcitonin and a parathyroid hormone analog, by genetic profiling analysis of multiple organs in primates. It is also believed that this is the first genetic profiling analysis that describes the molecular mechanisms of action of hormone-mediated bone remodeling by salmon calcitonin and PTS893. In this Example, it was found that both salmon calcitonin and PTS893 have modulatory effects on the genes that affect the direct, autocrine, paracrine, and endocrine regulation of mesenchymal cell functions, such as transforming growth factors betas (TGF). -ps), insulin-like growth factors (IGFs), bone morphogenetic proteins (BMPs), and vascular endothelial growth factor (VEGF). Both compounds also regulate the synthesis and degradation of extracellular matrix components. Salmon calcitonin also regulates the estrogen receptor and the steroidogenic factor, while PTS893 produces a strong increase over the nuclear receptors of the steroid / thyroid receptor family. Therefore, these data support the role of calcitonin as an anabolic agent. In addition, salmon calcitonin and PTS893, also had an influence on some aspects of extracellular matrix mineralization, due to changes in amelogenin, dentin, and pitufocytes of ectonucleotide. In addition, PTS893 showed an effect on the mediation of paracrine activation of differentiation and activity of osteoclasts, through the cytokine and the RANK ligand. No significant differences could be attributed in the profiling of gene expression to! fact of administering salmon calcitonin and PTS893 in combination, with respect to individual therapy. Therefore, the genetic profiling analysis of this example allowed us to reconstruct the pathways involved in the signal transduction of calcitonin and hormone for rati roid, triggered by the stimulation of the receptor bound with G-protein and its influence on the cycle cellular, as indicated by the changes observed in the cyclins. Animals. A two-week subcutaneous treatment was carried out with salmon calcitonin (sCT), PTS893, or a combination of the two, each of which was dissolved in phosphate-regulated serum (PBS) containing 9 percent autologous serum. A solvent was used as vehicle for the control group. The animals used in this analysis were cynomolgus monkeys (Macaca fascicularis), supplied by the Center de Recherches, Primatologiques, Port Louis, Mauritius. Two animals were used per group and sex. At the beginning of the treatment period, the animals were at least 24 months of age, with a body weight of approximately 3 kilograms. The animals were kept under standard conditions for the welfare of the animal. The animals were examined daily to determine mortality, feed intake, and for clinical observations. Body weight was recorded once a week. The dosages were 0 micrograms / animal / day (as the control), 50 micrograms / animal day of salmon calcitonin, and 5 micrograms / animal / day of PTS893. As shown later, the clinical observations and analysis, as well as the histopathological examinations carried out in this Example, showed that salmon calcitonin administered subcutaneously in a dose of 50 micrograms / animal / day, was well tolerated by cynomolgus monkeys. In vivo exams No significant histopathological changes were observed. No relevant changes were observed other than a reduction in body weight that was in the range of 8 to 12 percent in the group with salmon calcitonin. A reduction in feed intake was also observed, although it was not always consistent with the reduction in body weight. TABLE 1 Food Consumption - Males Control Salmoncalcitonin Day -6 |5 -4 -3 -2 -1 1 2 3 4 5 Animal no. W62503 50 75 50 75 100 75 75 75 50 100 100 Animal no. W62504 50 75 75 75 100 75 50 25 100 75 100 Day 6 7 8 9 10 11 12 13 14 Prom Animal no. W62503 75 100 100 100 100 75 75 25 70.8 Animal no. W62504 75 75 75 100 100 75 75 25 75 75.0 Both animals 72.9 PTS893 The animals given the salmon calcitonin showed a reduction in body weight that was in the range of between 8 and 12 percent, which can be attributed to a reduction in feed intake. An anorectic effect had previously been described for salmon calcitonin, which acts through the amylin receptors, Eiden S. et al., J. Physiol. 541 (pt3): 1041-1048 (2002); Lutz T. A. et al., Peptides 21 (2): 233-8 (2000). However, no signs of toxicity were observed here. The hormonal and lipid changes observed in this example are most likely related to a consequent metabolic adaptation. No relevant changes were observed in electrocardiograms (ECG) or blood pressure.
TABLE 2 Blood Pressure Animal Compound Sex Week 1 Week 2 Difference no. administered (mm Hg) (mm Hg) (mm Hg) W62501 Male Control 121 98 -23 W62501 Male Control 90 29 -61 W62502 Male Control 86 107 21 W62502 Male Control 26 34 8 W62503 Male Calcitonin 135 99 -36 Salmon W62503 Male Calcitonin 61 40 -21 Salmon W62504 Male Calcitonin 102 79 -23 Salmon W62504 Male Calcitonin 56 35 -21 Salmon W82505 Male PTS893 76 87 11 W82505 Male PTS893 18 22 4 W82506 Male PTS893 106 101 -5 W82506 Male PTS893 53 33 -20 W62551 Female Control 96 76 -20 W62551 Female Control 27 26 -1 W62552 Female Control 102 93 -9 W62552 Female Control 26 36 10 W62553 Female Calcitonin 98 82 -16 Salmon W62553 Female Calcitonin 50 25 -25 Salmon W62554 Female Calcitonin 92 44 -48 Salmon W62554 Female Calcitonin 26 30 4 Salmon W62555 Female PTS893 92 70 -22 W62555 Female PTS893 43 42 -1 W62556 Female PTS893 78 87 9 W62556 Female PTS893 24 28 4 Blood sampling. The animals were fasted during the night before the blood collection, but had free access to water. Blood samples were taken from a peripheral vein. Standard analyzes of hematology and clinical chemistry were carried out once during the pre-test, and at the end of the treatment period. Blood samples were collected from each animal at the same intervals that are described for clinical chemistry investigations. The serum samples were deep frozen (at approximately -80 ° C) until analysis, for the determination of hormones.
Clinical chemistry and hormone determinations.
A slight anemia was observed in all the study animals, including the controls. This was attributed to repeated blood sampling, and was not considered relevant.
TABLE 3 Hematology - Machos Control Animal no. W62501 W62502 Test Units d-6 d7 d13 d-6 d7 d13 WBC G / l 10.0 11.1 12.9 6.1 11.2 6.3 RBC T / l 7.3 6.5 6.4 6.8 6.5 6.2 HB g / di 12.9 11.9 11.7 13.1 12.3 11.9 PCV l / l 0.44 0.40 0.44 0.42 0.41 0.41 MCV fl 60 61 68 61 63 66 MCH pg 17.8 18.2 18.1 19.3 19.0 19.0 CHC g / di 29.8 29.6 26.8 31.5 30.1 28.9 PLAT G / l 316 371 266 458 500 547 NG / l 6.46 4.93 3.65 2.09 6.77 1.24 EG / l 0.01 0.14 0.20 0.10 0.10 0.10 BG / i 0.02 0.03 0.06 0.02 0.02 0.00 LG / l 3.05 5.45 8.44 3.60 3.65 4.51 MG / l 0.46 0.51 0.54 0.33 0.64 0.46 Salted Calcitonin d-6, d7, and d13 indicate day -6, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 3 Hematology - Males PTS893 d-6, d7, and d13 indicate day -6, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 4 Hematology - Females Control d-8, d7, and d13 indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 4 Hematology - Females Calcitonin Salmon PTS893 d-8, d7, and d13 Indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage. Among the standard clinical chemistry tests carried out, there were slight to moderate decreases in phosphorus and / or magnesium, and a moderate, marked decrease in triglycerides, in the groups given salmon calcitonin and PTS893 .
TABLE 5 Clinical Chemistry - Machos Control d-6, d7, and d 3 indicate day -6, day 7, and day 13, in relation to the day of the beginning of the dosage.
TABLE 5 Clinical Chemistry - Males Calcitonin Salmon d-6, d7, and d13 indicate day -6, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 5 Clinical Chemistry - Males PTS893 d-6, d7, and d13 indicate day -6, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 6 Clinical Chemistry - Females Control d-8, d7, and d13 indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 6 Clinical Chemistry - Females Calcitonin Salmon d-8, d7, and d13 indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 6 Clinical Chemistry - Females PTS893 d-8, d7, and d13 indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage.
No relevant changes were observed in the conventional urinalysis tests carried out.
TABLE 7 Urinalysis | Males Control Animal no. W62501 W62502 Test Units -6 -5 13 -6 -5 13 VOLUME mi 15 10 77 22 130 30 CREAT μ ???? /? 18000 17000 5460 7920 2480 5160 NTx n ECB - 9954 3425 - 1197 3167 9 CTx M9I - 21592 6810 - 2716 5323 9 D-PYR nmol / l 2345 1110 2904 1461 LDH IU / L 6.0 nd 8.0 8.0 NAG IU / 1 3.5 1.5 3.2 1.6 Na + mmol / l 163 43 87 77 K + mmol / l 258 67 125 75 Cl- mmol / l 132 43 52 59 Ca2 + mmol / l 5.15 16.80 15.95 15.50 LPHOS mmol / l 11.10 1.05 11.30 8.90 Mg2 + mmol / l 2.75 7.50 7.85 6.25 Na / Crea m / m 9.10 7.90 1 .00 14.90 K / Create m / mM 14.30 12.20 15.80 14.50 C / Crea mM / mM 7.40 7.90 6.50 11.40 Ca / Crea m / mM 0.29 3.08 2.01 3.00 Pho / Crea mM / mM 0.62 0.19 1.43 1.73 g / Create mM / m 0.20 1.40 1.00 1.20 LDH / creates lU / mM 0.33 nd 1.01 1.55 NAG / create lU / mM 0.19 0.28 0.40 0.31 NTx / Crea n E / mM 586 627 4830 614 CTx / Crea 1270 1247 10955 1032 Pyr / Crea nM / m 138 203 1171 283 d-6, d-5, and d 13 indicate day -6, day -5, and day 13, in relation to the day of the beginning of the dosage.
TABLE 7 Urinalysis - Males Calcitonin Salmon Animal no. W62503? W62504 Test Units -6 -5 13 -6 -5 13 VOLUME mi 62 38 68 37 10 54 CREAT μ ???? /? 4300 7840 4620 13600 17360 4400 NTx n ECB 6023 5186 16067 3790 CTx ug / i 11618 10088 26370 6130 D-PYR nmol / l 1733 1083 _ 5113 1476 LDH IU / L 9.0 7.0 13.0 17.0 NAG IU / 1 2.7 1.4 4.2 7.2 Na + mmol / l 22 14 119 15 K + mmol / l 65 78 134 76 Cl- mmol / l 10 55 64 68 Ca2 + mmol / l 0.90 18.25 3.70 23.40 LPHOS mmol / l 4.35 2.50 5.33 3.00 Mg2 + mmol / i 1.40 7.05 7.55 9.80 NaCrea mM / m 5.20 3.10 8.70 3.40 KCrea mM / m 15.10 16.90 9.90 17.20 C / Crea mM / mM 2.20 11.80 4.70 15.30 Ca / Crea mM / mM 0.21 3.95 0.27 5.32 Pho / Crea mM / m 1.01 0.54 0.39 0.68 Mg / Crea mM / mlM 0.30 1.59 0.60 2.20 LDH / creates lU / mM 2.09 1.52 0.96 3.86 NAG / create lU / mM 0.63 0.30 0.31 1.64 NTx / Create nME / mM 768 1123 926 861 CTx / Create μ9 / μ? T? 1482 2184 1519 1393 Pyr / Crea nM / m 221 234 295 336 d-6, d-5, and d13 indicate day -6, day -5, and day 13, in relation to the day of beginning of the dosage.
TABLE 7 Urinalysis | Males PTS893 Animal no. W62505 W62506 Test Units -6 -5 13 -6 -5 13 VOLUME mi 14 14 48 58 34 130 CREAT μ ???? /? 16160 16160 7840 9940 16120 3840 NTx nM BCE 5403 4871 8757 2102 CTx Mg / I - 11865 9365 - 2010 3705 8 D-PYR nmol / l 1660 1676 2278 782 LDH IU / L 7.0 14.0 9.0 19.0 NAG IU / 1 23.4 2.9 7.1 2.6 Na + mmol / l 174 111 59 35 K + mmol / l 86 107 125 69 Cl- mmol / l 22 117 50 48 Ca2 + mmol / l 5.10 7.55 3.50 13.10 LPHOS mmol / l 74.40 0.10 3.86 0.17 Mg2 + mmol / l 11.25 8.70 2.95 5.25 Na / Crea m / mM 10.80 14.10 6.00 9.10 K / Create mM / mM 5.30 13.60 12.60 17.90 C / Create mM / mM 1.40 15.00 5.00 12.60 Ca / Create mM / mM 0.32 0.96 0.35 3.41 d-6, d-5, and d13 indicate the di -6, the day -5, the 13th, in relation to the start day of dosing TABLE 8 Urinalysis - Females Control Animal no. W62551 W62552 Test Unit -8 -7 13 • 8 -7 13 VOLUME mi 21 21 43 18 53 53 CREAT Mmol / I 16420 16420 9560 14300 6700 5380 NTx n ECB 9248 7824 5053 4695 CTx pgi 19280 17916 12014 10557 D-PYR nmol / l 2500 2748 1397 2159 LDH IU / L 10.0 15.0 9.0 25.0 NAG IU / 1 19.2 4.2 10.3 3.5 Na + mmol / l 110 44 140 64 K + mmol / i 82 122 124 87 Cl-mmol / l 24 73 72 56 Ca2 + mmol / l 2.90 16.10 11.90 19.90 19.50 LPHOS mmol / l 88.2 7.7 20.3 3.5 g2 + mmol / l 2.35 7.20 9.00 5.45 Na / Crea mM / m 6.70 4.60 9.80 11.90 K / Crea m / m 5.00 12.80 8.70 16.20 C / Crea mM / m 1.50 7.60 5.10 10.50 Ca / Crea m / mM 0.18 1.68 0.83 3.63 P o / Create mM / mM 5.37 0.81 1.42 0.64 Mg / Crea m / mM 0.10 0.80 0.60 1.00 LDH / creates lU / mM 0.61 1.57 0.63 4.65 NAG / create lU / mM 1.17 0.44 0.72 0.65 NTx / Crea nME / mM 563 818 754 783 CTx / Crea MgMm 1174 1874 1793 1962 Pyr / Crea nlWmM 152 288 209 401 d-8, d-7, and d13 indicate day -8, day -7, and day 13, in relation to the day of the beginning of the dosage.
TABLE 8 Urinalysis - Females Calcitonin Salmon Animal no. W62553 W62554 Test Units -8 -7 13 |8 -7 13 VOLUME mi 11 58 67 32 14 49 CREAT μ ???? /? 10780 6920 4800 11260 1338 4200 0 NTx nM ECB 4624 3465 7393 2812 CTx MQ / i - 6983 5392 - 1341 5631 1 D-PYR nmol / l 2762 1644 2016 1110 LDH IU / L 14.0 6.0 6.0 36.0 NAG IU / 1 10.2 2.8 1.2 2.7 Na + mmol / l 98 40 156 32 K + mmol / l 104 53 172 57 Cl- mmol / l 31 63 156 65 Ca2 + mmol / l 3.00 17.55 3.50 12.70 LPHOS mmol / l 25.4 5.1 10.8 5.8 Mg2 + mmol / l 3.35 5.40 3.80 4.85 Na / Crea mM / m 9.10 8.30 13.90 7.60 K / Create mM / mM 9.60 11.10 15.20 13.50 C / Create mM / mM 2.90 13.20 13.80 15.40 Ca / Create imM / m 0.28 3.66 0.31 3.02 Pho / Crea mM / m 2.35 1.05 0.96 1.38 Mg / Crea mM / m 0.30 1.10 0.30 1.20 LDH / creates lU / mM 1.30 1.25 0.53 8.57 NAG / create lU / mM 0.95 0.58 0.11 0.64 NTx / Create nME / mM 668 722 553 670 CTx / Create 1009 1123 1002 1341 Pyr / Crea nM / mM 399 343 151 264 d-8, d-7, and d13 indicate day -8, day -7, and day 13, in relation to the day of the beginning of the dosage.
TABLE 8 Urinalysis - Females PTS893 Animal no. W62555 l W62556 Unit Test -8 -7 13 -8 -7 13 VOLUME mi 14 15 52 39 69 42 CREAT μ ???? /? 19160 18240 5620 14060 7600 8060 NTx nM BCE 10499 2514 4818 5679 CTx 21919 3813 8877 11236 D-PYR nmol / l 2963 1356 1377 2036 LDH IU / L 11.0 10.0 18.0 9.0 NAG IU / 1 0.5 1.2 5.9 5.1 Na + mmol / l 145 71 118 146 K + mmo! / 1 302 50 164 70 Cl-mmol / l 119 101 53 133 Ca2 + mmol / l 11.50 20.05 6.60 12.35 LPHOS mmo! / L 0.2 0.1 7.6 2.9 Mg2 + mmol / l 7.35 6.90 4.00 5.90 NaCrea mM / mM 7.60 12.60 8.40 18.10 K / Create mM / m 15.80 26.80 11.70 8.60 C / Crea mM / mM 6.20 18.00 3.70 16.50 Ca / Crea mM / m 0.60 3.57 0.47 1.53 Pho / Crea mM / mM 0.01 0.02 0.54 0.36 Mg / Crea mM / mM 0.40 1.20 0.30 0.70 LDH / creates lU / mM 0.57 1.78 1.28 1.12 NAG / create lU / mM 0.03 0.21 0.42 0.63 NTx / Crea n E / m 576 447 634 705 CTx / Crea Mg / pm 1202 679 1168 1394 Pyr / Create nM / m 163 241 181 253 d-8, d-7, and d13 indicate day -8, day -7, and day 13, in relation to the day of beginning of the dosage. The salmon calcitonin group showed moderate decreases in serum somatomedin (S.MED., See Tables 9 and 10).
TABLE 9 Hormones - Machos Control d-6, d7, and d13 indicate day -6, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 9 Hormones - Machos Control Animal no. W62503 W6250 ¿l Test Units d-6 d7 d13 d-6 d7 d13 ACTH pg / ml 98 87 87 115 78 73 CORTISOL nmol / l 2316 979 1611 1578 1523 1709 ALDOST pg / ml 983 1058 819 465 987 977 INSULINA mU / l 13.0 14.0 17.0 4.0 10.0 22.0 GLUCAG pg / ml 905 247 428 869 218 503 C-PEPTI ng / ml n / a 1.70 1.80 n / a 1.20 2.30 GASTRINE pg / ml n / a 83 88 n / a 128 136 d-6, d7, and d13 Indicate day -6, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 9 Hormones - Males PTS893 6, d7, and d 13 indicate the day -6 on day 7, and day 13, ation with the day of beginning of the dosage TABLE 10 Hormones - Females Control G5 Animal no. W62551 W62552 Test Units d-8 d7 d13 d-8 d7 d13 ACTH pg / ml 146 276 121 58 60 101 CORTISOL nmol / l 1983 1546 827 1894 837 818 0 ALDOST pg / ml 244 953 312 149 90 199 INSULININ mU / l 8.0 12.0 7.0 2.0 29.0 21.0 GLUCAG pg / ml 729 779 583 818 507 514 C-PEPTI ng / ml n / a 2.40 1.40 n / a 3.30 2.30 GASTRINE pg / ml n / a 84 102 n / a 90 92 5 d-8, d7, and d13 indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage.
TABLE 10 Hormones - Females Calcitonin Salmon , d7, and d 13 indicate day -8 on day 7, and day 13, ation with the day of beginning of the dosage TABLE 10 Hormones - Females PTS893 Animal no. W62555 \ V62556 Test Units d-8 d7 d13 d-8 d7 d13 ACTH pg / ml 109 104 110 95 132 126 CORT1SOL nmol / l 1482 1331 917 1532 1253 1375 ALDOST pg / ml 314 217 330 210 228 226 INSULINA mU / l 1.0 22.0 19.0 15,0 30.0 22.0 GLUCAG pg / ml 711 591 657 696 437 380 C-PEPTI ng / ml n / a 3.00 2.40 n / a 3.80 3.50 GASTRINE pg / ml n / a 83 82 n / a 96 91 d-8, d7, and d13 indicate day -8, day 7, and day 13, in relation to the day of beginning of the dosage.
Sampling of tissue. The animals were sacrificed by deep anesthesia induced by an intravenous injection of Pentothal®, followed by exsanguinations. All relevant tissues were sampled for histopathology and the expression of gene expression. The following tissue samples were processed for analysis: liver, kidney, pituitary, muscle, bone, duodenum, spleen, and trachea. Samples for histopathology were fixed in 10 percent formalin regulated with phosphate. The bone demineralization was carried out with 10 percent formic acid. The tissue samples were embedded in Paraplast®, and sectioned at 4 microns, to stain with hematoxylin and eosin. Samples for profiling of gene expression were rapidly frozen in liquid nitrogen immediately after cutting, stored on dry ice, and subsequently in a deep freezer at about -80 ° C until further use. All tissues selected for profiling of gene expression were examined histopathologically. Histopathology The histopathological examination of the tissues selected for the genetic profiling analysis showed a normal spectrum of incidental lesions that, in terms of severity and distribution of the lesions, were not different from the controls in all the treatment groups. A slightly higher incidence of inflammatory and regenerative changes was observed in the kidneys of females given salmon calcitonin. These changes were not considered relevant, because there are no records of kidney toxicity after 40 years of therapeutic use of calcitonin. The bone sections were stained for osteonectin, osteopontin, and osteocalcin, and were evaluated histopathologically. The histomorphometry of the bone tissue was carried out with respect to the parameters for bone resorption and synthesis (osteoid formation). The staining with osteonectin, osteopontin, and osteocalcin of the tibia showed no difference between groups 1 (control) and two (salmon calcitonin). The osteonectin exhibited a greater aggravation and deterioration of the epiphyseal growth plate of animal number 2553, due to a severe pathological condition unrelated to treatment (severe subacute epiphysis). The histomorphometry of the bone tissue was carried out to determine the parameters related to bone resorption and bone synthesis (osteoid formation). The results (see Tables 11 and 12) showed that salmon calcitonin increased trabecular volume and thickness by approximately 17 percent in the tibia, but not in the vertebrae. PTS893 reduced cortical thickness (18 percent) and increased cortical porosity (54 percent) in the tibia (T), but not in the vertebrae (V). In contrast, PTS893 induced an increase in osteoid volume (37 percent in the tibia, 213 percent in the vertebrae), and in the osteoid surface (49 percent in the tibia, 37 percent in the vertebrae), as well as an increase in the surface of osteoblasts (40 percent in the tibia, 24 percent in the vertebrae), both in the tibia and in the vertebrae, respectively.
TABLE 11 Histomorf ometry of the Tibia (Average of Males and H em bras) average 25.83 126.88 2.01 391.68 2.02 1009.14 34.97 6.65 4.95 15.66 SD 7.17 10.68 0.43 144.81 0.33 124.65 5.56 3.46 1.93 6.21 PTS893 19.69 129.22 1.52 526.99 2.76 1022.62 54.84 .24 4.62 16.16 16. 65 93.20 1.79 466.69 2.94 893.43 43.57 9.61 4.76 21.25 25. 74 120.52 2.13 347.63 2.94 950.33 43.63 8.14 4.21 18.46 24. 78 126.07 1.97 382.61 2.95 939.53 54.97 9.95 2.85 25.25 average 21.72 117.25 1.85 430.98 2.90 951.48 49.25 9.74 4.11 20.28 SD 4.30 16.43 0.26 81.20 0.09 53.46 6.53 1.28 0.87 3.91 sCT: Calcitonin from salmon; SD: Standard deviation. BV / TV, trabecular bone volume; Tb. Th. Trabecular thickness; Tb. N. Trabecular number; Tb. Sp. Trabecular separation; Ct. By. Cortical porosity; Ct. Th. Cortical thickness; OS / BS, osteoid surface; OV / BV, osteoid volume; ES / BS, eroded surface; Obs / BS, superficial osteoblasts.
TABLE 12 H istom or rfom etry of the Vertebrae (Average of Males and H em bras) average 19.75 144.72 1.39 598.07 0.71 637.20 18.47 1.45 6.36 7.74 SD 2.82 25.07 0.28 138.62 0.30 193.78 5.15 0.45 3.34 5.65 sCT 17.32 113.29 1.53 540.84 1.70 705.10 3.95 0.46 11.60 3.21 19. 33 144.31 1.34 602.15 1.18 810.09 5.82 0.86 2.55 3.97 20.11 118.49 1.70 470.71 1.18 576.42 11.48 1.43 4.93 6.81 19. 46 123.71 1.57 511.96 0.12 907.16 4.91 0.32 3.47 1.23 average 19.06 124.95 1.53 531.42 1.05 749.69 6.54 0.77 5.64 3.80 SD 1.21 13.59 0.15 55.24 0.66 141.96 3.38 0.50 4.09 2.31 PTS893 15.15 105.46 1.44 590.67 1.49 707.43 18.84 3.24 9.31 10.36 20. 23 118.79 1.70 468.39 1.45 629.35 41.28 8.42 2.30 9.07 23.56 134.66 1.75 436.79 0.41 740.87 23.65 3.49 2.55 10.47 24. 86 134.82 1.84 407.56 0.92 624.35 17.66 2.66 3.96 8.33 average 20.95 123.43 1.68 475.85 1.07 675.50 25.36 4.45 4.53 9.56 SD 4.33 14.15 0.17 80.47 0.51 57.85 10.93 2.67 3.27 1.04 sCT: Calcitonin from salmon; SD: Standard deviation. BV / TV, trabecular bone volume; Tb. Th. Trabecular thickness; Tb. N. Trabecular number; Tb. Sp. Trabecular separation; Ct. By. Cortical porosity; Ct. Th. Cortical thickness; OS / BS, osteoid surface; OV / BV, osteoid volume; ES / BS, eroded surface; Obs / BS, superficial osteoblasts.
The histomorphometry showed inconsistent results between the tibia bone and the vertebrae, except for an increase in osteoid synthesis induced by PTS893. This effect is well documented for the parathyroid hormone, when administered in a discontinuous manner. Extraction and purification of RNA. A set of tissues was selected for the profiling of gene expression. This set included samples of kidney, bone, muscle, duodenum, pituitary, and liver. In particular, the diaphysical bone of the femur and tibia was processed for the profiling of the gene expression. Briefly stated, the total RNA was obtained by extraction with guanidinium-phenol-chloroform thiocyanate thiocyanate (Trizol®, Invitrogen Life Technologies, Carlsbad, Calif. EUA) of each section of frozen tissue, and then the total RNA was purified on an affinity resin (RNeasy®, Qiagen) according to the manufacturer's instructions. Total RNA was quantified by absorbance to? = 260 nanometers (A260nm), and the purity was estimated by the proportion of A260nm / A280nm. The integrity of the RNA molecules was confirmed by non-denaturing agarose gel electrophoresis. RNA was stored at approximately -80 ° C until analysis. A portion of each individual RNA sample was saved for analysis of critical genes by means of real-time polymerase chain reaction. Hybridization assay. Transcription profiling was done by means of arrays of GeneChip® expression probes in Genomics Factory EU laboratories, as recommended by the manufacturer of the GeneChip® system (GeneChip Expression Analysis Technical Manual, Affymetrix Inc., Santa Clara, Calif. USA). HG-U95Av2 GeneChip® expression probe arrays were used (Affymetrix, Santa Clara Calif. USA). The double-stranded cDNA was synthesized with an initial amount of about 5 micrograms of full length total RNA, using the Superscript Choice System (Invitrogen Life Technologies) in the presence of an oligonucleotide primer of T7- (dT) 24 DNA. Following the synthesis, the cDNA was purified by extraction with phenol / chloroform / aminosol, and ethanol precipitation. The purified cDNA was then transcribed in vitro, using the BioArray® high-throughput RNA Transcription Marking Kit (ENZO) in the presence of biotinylated ribonucleotides of biotin-labeled cRNA. The labeled cRNA was then purified on an affinity resin (Rneasy®, Qiagen), quantified, and fragmented. An amount of about 10 micrograms of labeled cRNA was hybridized for about 16 hours at 45 ° C, to an array of expression probe. Then the array was washed and stained twice with streptavidin-phycoerythrin (Molecular Probes), using the GeneChips Fluidics 400 workstation (Affymetrix). The array was then scanned twice using a confocal laser scanner (GeneArray® Scanner, Agilent), resulting in a scanned image. This "file- .data" was processed using the Micro Array Analysis Suite version 4 (MAS4) program (Affymetrix) in a "file-.cel". The "cel- file" was captured and loaded into the Affymetrix GeneChip Laboratory Information Management System (LIMS). The LIMS database is connected to a Sun Solaris UNIX server through a network file system that allows the average intensities for all probe cells (CEL file) to be downloaded into an Oracle database. The raw data was converted to the expression levels using an "objective intensity" of 150. The numerical values displayed are the weighted averages of the signal intensities of the pairs of probes comprised in a set of probes for a given transcription sequence ( PromDif value). The quality of the data was verified and loaded into the GeneSpring® software versions 4.2.4 and 5 (Silicon Genetics, USA) for analysis. Analysis of data. The data analysis was carried out with the software package of Silicon Genetics, GeneSpring, version 4.2.1 and 5. The average difference values below 20 were set to 20. Different filtering and accumulation tools were used in these programs , to explore the data sets and to identify the changes in the level of transcription that inform about altered cellular and tissue functions, and that can be used to establish working hypotheses about the modes of action of the compound. The threshold interval for consideration of up or down regulation was determined within the context of the biological interpretation of the example. The information content of these data sets is a set of numerical changes and biological information. The decision to consider a specific relevant gene was based on a set of numerical changes identified by the comparative and statistical algorithms, and the relationship with other modulated genes that point to a common biological issue. The weight of that relationship was evaluated by the analyst through a review of the relevant scientific literature. The increase and decrease reported here refer to the abundance of the transcript, unless specifically reported. Perfilation of gene expression. The analysis of comparative genetic profiling of multiple organs was carried out in the group in which salmon calcitonin was administered at 50 micrograms / year / day / bad. The organs selected for the analysis were liver, kidney, pituitary, skeletal muscle, bone, duodenum, spleen, and trachea.
TABLE 13 Profiling Genetic Expression of Multiple Organs of Salmon Calcitonin GenCip® Gene Coding Bone Expression Probe Set Identifier 36611 at acid phosphatase 1, isoform a or Riñn -1.33 -1.33 32714 s at 1 activin A type II receptor type -1.62 -1.83 í Hdgao 39314_at activin receptor precursor A -1.12 1.41 -4.15 type II B or Mlscuo 35915 at activin beta-C chain -1.21 -2.41 -1.67 Piittiuara 36621 at glycoprotein alfa-2-HS 1.33 1.53 1.12 34588 i at amelogenin -1.61 á Trquea 37747 at annexation V -1.30 1.87 -2.58 40376 at the arylsulfatase precursor E -1.59 39326 at ATPase H (+) - vacuolar -1.57 -2.80 -1.62 38814 at ATPase H (+) - vacuolar subunit 1.22 33741_at ATPase, transport of H +, 1.23 -1.50 lysosomal 33033_at ATPase, transport of H +, -1.29 -3.19 -1.43 1.23 lysosomal 38814_at ATPase, transport of H +, 1.30 -1.28 1.14 lysosomal 38126 at biqlicano 1.75 -1.61 39407 at bone morphogenetic protein 1 -1.20 -1.55 31399 at bone morphogenetic protein 10 1.44 1.45 -1.31 -1.77 1113 at bone morphogenetic protein 2A -1.12 2.63 1.29 1831 at bone morphogenetic protein 5 -1.43 1.39 1.40 1733_at precursor morphogenetic protein -1.37 -1.17 -1.64 -1.27 -1.27 -1.1 bone 6 34500_at calcium binding protein 1 2.31 1.21 (calbrain) 31670_s_at protein-dependent kinase 1.17 1.57 -1.28 1.60 calcium / calmodulin (CaM kinase) II gamma 1751 g at calreticulin -4.03 -1.60 1.67 32067_at element modulator 1.39 -1.24 -1.50 responds to cAMP (CRE) 39241 at carbonic anhydrase I -2.68 1.18 -1.69 40095 at carbonic anhydrase II -1.69 40163_r_at precursor of matrix protein 2.36 5.61 oligomeric cartilage 128 at cathepsin k 1.18 1.35 -2.33 129_g_at cathepsin k 1.20 -1.54 1.17 -1.28 38466 at cathepsin k 1.27 1.40 -1.19 39333 at collagen, type IV, alpha 1 -1.49 39925 at collagen, type IX, alpha 2 -2.38 -1.36 38420 at collagen, type V, alpha 2 -1.29 -1.18 -1.11 -1.10 41351 at collagen, type VI, alpha 1 -2.29 -1.27 -1.50 41350_at collagen precursor, type VI, alpha -3.55 1 35168 f at collagen, type XVI, alpha 1 -1.59 35169 at collagen, type XVI, alpha 1 -1.18 39632_at collagenase 3 (metalloproteinase of 1.20 matrix 13) 36638_at tissue growth factor -2.11 connective 40697 at cyclin A2 -1.60 34736 at cyclin B1 -2.83 36650 at cyclin D2 1.21 35249 at cyclin E2 -2.95 1206 at kinase 5 cyclin dependent 1.56 -1.54 799_at kinase 5 dependent cyclin, 1.32 regulatory subunit 1 (p35) 41546 at kinase 6 cyclin dependent 1.15 1.52 1.34 2031_s_at inhibitor 1A of kinase 1.95 dependent on cyclin (p21, Cip1) 35816 at cystatin B (stefin B) 1.57 806 at kinase inducible by cytokine 1.20 1.35 | 40049_at protein kinase 1 associated with -1.47 -1.29 death 33903_at protein kinase 3 associated with -1.22 death 34029_at acid phosphoprotein 1 matrix 1.65 dentin (D P1) 40186 at double specificity phosphatase 9 1.59 37996_s_at myotonic protein kinase of 1.25 -1.50 dystrophy 342_at pyrophosphatase / phosphodiesterase of 1.45 ectonucleotide 1 343_s_at pyrophosphatase / phosphodiesterase of 1.1 1 -1.42 ectonucleotide 1 33602_at endothelial differentiation, precursor 1.15 2.24 -1.66 of receptor 6 coupled with G protein 1442 at estrogen receptor 1.47 1.23 1.60 33670 at estrogen receptor 1.30 1487_at protein related to receptor 1.11 -1.52 1.24 estrogen 38882_r_at protein of B frame responding 1.22 -1.51 to estrogen (EBBP) 39945_at activation protein of -1.27 -1.48 -1.32 fibroblasts 996 at growth factor of 1.17 -1.41 41280_r_at MAPK8IP1: protein 1 of -1.31 1.92 1.58 interaction with protein kinase 8 activated by mitogen 2004 at MEK kinase 1.13 -1.62 1.16 1509 at metalloproteinase -1.42 -1.11 -1.23 -1.18 976_s_at kinase 1 of activated protein by -1.61 mitogen 34006_s_at kinase 8 of activated protein by 1.32 mitogen 1844_s_at kinase 1 protein kinase -1.60 1.15 mitogen-activated kinase 35694_at kinase 4 kinase 1.26 mitogen-activated protein kinase 1469_at protein kinase 2 activated by 113 -1.30 1.16 protein kinase mitogen-activated protein 1637_at kinase 3 of protein activated by 1.11 1.34 protein kinase activated by mitogen 37565_at MMD: associated with differentiation 1.28 -2.48 -1.28 of monocytes to macrophages 38307 at neurocondrina 2.80 -1.39 39144_at nuclear factor T-cells 2.72 1.42 -1.70 activated, cytoplasmic, (Drosophila) phosphodiesterase E3) 38921 at phosphodiesterase IB 1.52 1.42 1.12 31699_at 3-kinase phosphoinositide 1.56 -1.56 dependent on calmodulin 36287_at 3-kinase phosphoinositide, 1.31 catalytic, gamma polypeptide 35665 at 3-kinase phosphoinositide, class 3 -1.11 1.21 364 s at phospholipase Cb3 1.22 901_g at phospholipase C, beta 4 -1.20 1.41 -1.55 1293 s at phospholipase D -1.26 38023_at transfer protein 2.25 1.33 1.55 1.71 phosphatidyl-inositol 38269 at kinase D2 protein PKD2 1.34 32306 g at pre-pro-collagen type I, alpha-2 1.19 -1.38 -1.75 -1.31 35473 at pre-pro-collagen type I, alpha 1 -2.72 -1.37 -3.94 -2.70 32307 s at pro-collagen 1.13 -1.26 -2.44 -1.56 -1.82 37605 at pro-collagen alpha 1, type II -1.84 -1.61 36184_at pro-collagen 5-dioxygenase-2.52 -2.15 -1.30 lysine 37037_at pro-collagen-proline, 4- 1.87 1.46 -1.67 1.29 2-oxoglutarate dioxygenase (proline 4-hydroxylase), alpha polypeptide I 37633_s_at endometrial protein associated with 2.00 progestogen (placental protein 33679 f at tubulin beta 2 -1.31 1.45 709 ai tubulin beta 3 -1.18 -1.35 1.20 471 f at tubulin beta 4 -1.38 1.50 39399 at tubulin beta, cofactor D -1.85 -4.69 32098_at alpha-2 chain precursor of -3.79 type VI collagen 1651_at ubiquitin-carrier protein -3.74 E2-C 1953_at endothelial growth factor 1.40 vascular 36101_s_at endothelial growth factor 1.45 vascular 37268_at endothelial growth factor -1.58 vascular B 36140 at protein- 1 of frame link Y 2.30 1.86 2.36 -2.72 In addition, the effect of PTS893 on bone was evaluated.
TABLE 14 Analysis of Genetic Profiling of Salmon Calcitonin and PTS893 in Bone Identifier Times of times of set increase of probes with increase of coding gene expression Calcitonin with GeneChip® of salmon PTS893 38909_at 1-alpha-hydroxylase of 25-hydroxyvitamin -1.14 D3 32714 s at the activin A receptor type I l-type 1 -1.62 35915 at the beta-C chain of activin -1.21 39279 at the activin receptor type II 1.24 39383 at adenylate cyclase 6, isoform at -1.22 38965 at agrecano 1 2.03 39206 s at agrecano 1 1.41 36621 at glycoprotein alpha-2-HS 1.33 34589 f at amelogenlna 1.10 -3.10 39326 at ATPase h (+) vacuolar -1.57 -1.19 38814 at ATPase H (+) vacuolar 1.22 33741 at ATPase, transport of H +, lysosomal 1.23 33033_at ATPase, transport of H +, -1.29 -1.17 lysosomal 40328 at transcription factor bHLH 2.57 38466 at cathepsin k 1.27 40718 at cathepsin w -1.31 32833 at kinase type CDC 1 1.63 646 s at kinase type CDC 2, isoform hclk2 / 139 1.19 34763_at proteoglycan 6 sulfate -1.18 chondroitin 598 at collagen type i I, alpha -1.38 -1.19 32488 at collagen type III, alpha 1 -1.41 38952 s at collagen type IV, alpha 2 1.23 1.44 35379 at collagen type IX, alpha 1 -2.22 34802 at collagen type VI, alpha-2 (AA 570-998) -1.37 38566 at collagen type X, alpha 1 1.67 37892 at collagen type XI, alpha 1 1.24 1.18 1026 s at collagen type XI, alpha 2 -1.20 1027 at collagen type XI, alpha 2 1.11 39632_at collagenase 3 (metalloproteinase from 1.20 matrix 13) 36638_at tissue growth factor -1.32 connective 1943 at cyclin A -1.74 40697 at cyclin A2 -1.60 -1.39 34736 at cyclin B -2.83 39251 at cyclin C -2.03 1983 at cyclin D2 -1.28 36650 at cyclin D2 1.21 35249 at cyclin E2 -2.95 1649 at protein interaction with cicJina G1 1.31 1913 at cyclin G2 -1.29 160024_at cyclin-dependent kinase (type 1.53 CDC2) 10 PISSLRE 1942 s at cyclin-dependent kinase 4 -1.22 1206 at cyclin 5 dependent kinase 1.56 40549 at cyclin-dependent kinase 5 -1.40 799_at cyclin 5 dependent kinase, 1.32 regulatory subunit 1 (p35) 41546 at cyclin 6 dependent kinase 1.15 2031_s_at kinase inhibitor 1A dependent on 1.95 cyclin A (p21), Cip1) 1787_at kinase inhibitor 1 C dependent on 1-18 cyclin 38673_s_at kinase inhibitor 1 C dependent on 1.13 cyclin 39545_at inhibitor 1C kinase dependent on 1.24 cyclin 1797_at inhibitor 2D kinase dependent on -1.21 cyclin (p19, inhibits CDK4) 35816 at cystatin B (stefin B) 1.57 806 at cytokine-inducible kinase 1.20 40049_at protein kinase 1 associated with -1.30 death 33903_at protein kinase 3 associated with -1.22 -7.73 death 34029_at acid phosphoprotein 1 matrix 1.65 dentin (DMP1 ) 38059_g_at dermatopontina 1.72 343_s_at pyrophosphatase / phosphodiesterase 1.11 ectonucleotide 1 342_at pyrophosphatase / phosphodiesterase 1.45 ectonucleotide 1 1442 at the estrogen receptor 1.47 33670 at the estrogen receptor 1.30 1487_at 1.11 estrogen receptor-related protein 38882_r_at the B-frame protein that responds to 1.22 estrogen (EBBP) 38902_r_at Table B protein that responds to 1.23 estrogen (EBBP) 39945 at fibroblast activation protein -1.27 424_s_at growth factor receptor -1.17 fibroblasts 466 at general transcription factor II 1.34 1102 s at alpha glucocorticoid receptor 1.43 33510 s at the glutamate receptor, metabotropic 1 1.26 1.23 (somatomedin C) 1232_s_at factor binding protein -1.31 insulin-like growth 40422_at protein binding factor 2 -1.27 insulin-like growth 1586_at protein binding factor 3 1.45 growth insulin type 37319_at link protein 3 factor 2.17 insulin-like growth 1737_s_at link protein 4 factor 1.13 insulin-like growth 41420_at protein binding factor 1.18 insulin-like growth 1396_at protein binding factor 1.62 insulin-like growth 1678_g_at protein binding factor 1.44 insulin-like growth 38650_at protein binding factor 1.53 insulin-like growth 1741_s_at protein binding factor 2 -2.49 -2.11 growth type insulin 1464_at precursor growth factor type 1.18 insulin II 1591_s_at precursor of growth factor type 1.41 1.31 insulin II eno 41280_r_at MAP 8IP1: interaction protein -1.31 -1.31 with protein kinase 8 activated by mitogen 1509 at metalloproteinase -1.42 976_s_at protein kinase 1 activated by -1.61 1.12 mitogen 34006_s_at kinase 8 protein activated by 1.32 mitogen 1439_s_at kinase 2 protein activated by 1.78 mitogen-activated protein kinase 37565_at MMD: associated with differentiation from 1.28 1.30 monocytes to macrophages 38369_at primary response gene -1.10 myeloid differentiation (88) 1052 s at protein NF-IL6-beta 1.30 36472 at interaction with N-myc and STAT -1.35 38354 at nuclear factor NF-IL6 (AA 1-345) 1.92 33106_at nuclear receptor subfamily LXR-alpha 3.29 nuclear orphan receptor 1, group H, member 3 33381 at coactivator nuclear receptor 1.11 279_at nuclear receptor subfamily 4, group 2.30 A, member 1 | inositol, isoform C (-1) 751 at phosphatidylinositol-glycan-class C (PIG-C) 1.14 -1.25 666 at phosphodiesterase 4A, specific for cAMP 1.33 1.30 38526 at phosphodiesterase 4D, specific for cAMP 1.30 3.53 38921_at phosphodiesterase IB, dependent on 1.52 calmodulin 38944_at phosphodiesterase IB, dependent on 1.17 calmodulin 32029_at kinase-1 protein-dependent 1.16 phosphoinositide (3) 31699 at 3-phosphoinositide kinase 1.56 1.16 1085 s at phospholipase C -1.14 364 s at phospholipase C b3 1.22 901. g_at phospholipase C, beta 4 -1.20 1293 s at phospholipase D -1.26 32306_g_at pre-pro-collagen type I, alpha 2 1.19 35473 at pre-pro-collagen type I, alpha 1 -2.72 38951 at protein kinase C PRKCQ, tit 1.43 32307 s at pro-collagen 1.13 34494 at pro-collagen proteinase l-N 1.92 37605 at procollagen type II, alpha 1 1.91 36109 at prolidasa (imidodipeptidase) PEPD -2.55 1884_s_at cell nuclear antigen -1.85 proliferating 34390_at alpha (II) subunit of proline 1.19 proline 37037_at alpha subunit of 4-hydroxylase 1.20 prolyl 36666 at 4-hydroxylase prolyl beta 1.95 36533 at prostacycline synthase 1.20 718 at protease, serine, 11 (IGF binding) -1.30 7 9_g_at protease, serine, 11 (IGF binding) -1.43 385_at proteasome subunit (prosoma, 1.36 macropain), beta type, 10 39183 at protein kinase 1 PCTAIRE -1.17 37698_at protein kinase A (PRKA), anchor 1.29 protein 1 3971 1_at substrate 80K-H protein kinase 1.13 protein 39161 protein kinase Njmu-R1 1.21 35348_at protein kinase, activated by AMP, 2.10 non-catalytic beta 1 subunit 36359_at protein kinase, dependent on 1.39 cAMP, catalytic, gamma 546_at protein kinase, dependent on 1.14 cAMP, catalytic, protein inhibitor alpha 227_g_at protein, dependent on 1.18 cAMP, regulatory, type I, alpha 41768 protein kinase, dependent on 1.15 32548 at inactive progesterone receptor -1.33 1953_at endothelial growth factor 1.40 1.20 vascular 36101_s_at endothelial growth factor 1.45 1.44 vascular 36140 at protein-1 linkage box and 2.30 5.49 numbers (-) = down regulated. numbers (+) = regulated upwards.
Real-time polymerase chain reaction. Based on data from the DNA microarray, a set of transcripts was selected for quantitative analysis by real-time polymerase chain reaction (RT-PCR). Briefly stated, the method exploits the green SyBr dye, which is inserted into the double-stranded DNA. The accumulation of the products of the polymerase chain reaction is detected directly by monitoring the increase in fluorescence of the green SyBr dye. the reactions are characterized by the point of time during cycling when the amplification of a polymerase chain reaction product is first detected, rather than the amount of the polymerase chain reaction product accumulated after a number Fixed cycles. The higher the initial copy number of the nucleic acid target, the sooner a significant increase in fluorescence will be observed. From each RNA sample, the cDNA was made using the Applied Biosystems parts kit (Applied Biosystems # N808-0234), following the manufacturer's recommendation. The polymerase chain reaction mixture was prepared using the SyBr Universal Green Polymerase Chain Reaction Master Mix (Applied Biosystems # 4309155) as follows: 5 microliters of cDNA template, 400 nM of each primer, triphosphates of 0.2 mM deoxynucleotide, 1 mM MgCl2, and 0.5 units of Taq DNA polymerase, 5 microliters of SyBr green polymerase chain reaction regulator, and water without RNSA to a final volume of 50 microliters. The polymerase chain reaction was carried out using the Abl Prism 7700 Sequence Detection System, after one step at 95 ° C for 10 minutes, and the step cycle program was carried out for a total of 40 cycles as follows: 95 ° C for 30 seconds, 60 ° C for 1 minute. A negative control was included: polymerase chain reaction mixture with water instead of the cDNA sample. The initial template concentration was determined based on the threshold cycle. The threshold cycle is the cycle of the polymerase chain reaction where the fluorescence was first detected on the background, and it has been shown to be inversely proportional to the number of objective copies present in the sample. The quantification was carried out by calculating the unknown objective concentration in relation to an absolute standard, and by normalization with a validated endogenous control, such as a maintenance gene (β-actin). The results are presented as a percentage of the control, once the proportion between the numbers of molecules for the gene of interest divided by the number of molecules for β-actin has been calculated. Based on data from the DNA microarray, the following set of transcripts was selected for quantitative analysis by RT-PCR: CD44 adhesion receptor, ang iopoietin, bone morphogenetic protein 5, carbonic anhydrase II, cartilage oiigomeric matrix protein, cathepsin K, osteopontin, type I collagen pre-pro-alpha-2, Spi-B, and Y-frame binding protein.
TABLE 15 d ation in Polymerase Chain in Real Time: not applicable.
The real-time polymerase chain reaction confirmed in most cases the changes observed in the genetic profiling analysis, as was the case for the morphogenetic protein 5, carbonic anhydrase II, cathepsin K, oligomeric matrix protein of cartilage, type I collagen pre-pro-alpha-2, SpiB, and Y-frame binding protein. However, no changes were detected in the level of expression of the adhesion receptor CD44, angiopoietin 1, and osteopontin. Analysis. It is known that calcitonin exerts an effect on the differentiation, survival, and resorption activity of osteoclasts, resulting in decreased osteoclastic activity. Pondel Intl. J. Exp. Pathol. 81 (6): 405-22 (2000). These effects could be reconstructed by genetic profiling of multiple organs (Table 16).
TABLE 16 Effects on Osteoclasts osteoclast resorption activity) Bone resorption ATP-loops H + ALL B by osteoclasts Anhydrase B, L, carbonic P 1, 11 Cathepsin K ALL ODF / OPGL: B osteoproterenin ligand ALL Tubulin mobility osteoclasts Protein PAK4 B, M, P Profiling gene expression of multiple organs in animals treated with salmon calcitonin. The organs where the changes in the expression were seen are exhibited. B = bone; K = kidney; M = muscle; P = pituitary; L = liver, T = trachea. Salmon calcitonin appears to exert a paracrine regulation of the osteoclast resorption activity, through the regulation of cystatin expression in the osteoblast, as shown in Table 17.
TABLE 17 Genetic Expression Profiling: Osteoclast Function Probe Set Identifier Times of Controls sCT of Expression Gen Coding Average Average Change GeneChip® 40729_s_at ATPase, transport of 204 327 1.6 H +, lysosomal (vacuolar proton pump), G subunit, isoform 2 37367_at ATPase, transport of 272 328 1.2 H +, lysosomal, 31kDa, V1 subunit E, isoform 1 40568_at ATPase, transport of 938 1132 1.21 H +, lysosomal, 56 / 58kDa, V1 subunit E, isoform 2 39241 at carbonic anhydrase I 1266 441 -2.87 128 at cathepsin K (picnodisostosis) 5690 7821 1.37 129 g at cathepsin K (picnodisostosis) 5036 6757 1.34 38466 at cathepsin K (picnodisostosis) 5494 7267 1.32 36611 at acid phosphatase 1, soluble 254 331 1.3 PU.1 is involved in the initial stages of osteoclastogenesis. Tondravi. M. et al., Nature 386 (6620): 81-4 (1997). CSF- is imperative for the maturation of macrophages; it binds to its c-5 fms receptor on early osteoclast precursors, providing the signals required for its survival and proliferation. Teitelbaum SL, Science 289 (5484): 1504-1508 (2000). It is interesting that PTS893 also regulates the 0 genes involved in the differentiation and survival of osteoclasts, SPI 1, CSF-1, and MMD. This regulation of osteoclasts has not been described previously. It was shown that salmon calcitonin regulates the expression of the gene encoding osteoclast stimulating factor (OSF), which is an intracellular protein produced by osteoclasts, which indirectly induces osteoclast formation and bone resorption. Reddy S. et al., J. Cell Physiol. 177 (4): 636-45 (1998). This would imply an autocrine effect of salmon calcitonin on or regulation of osteoclast function, which is described here for the first time. In addition, salmon calcitonin appears to exert a paracrine regulation of the osteoclast resorption activity, through the regulation of cystatin expression in the osteoblast. Carbonic anhydrase I, II, ATPase H +, and cathepsin K, are the main effects to dissolve bone mineral degradation and matrix. Blair H. C. et al., Biochem. (2002). The regulation of tubulins and PAK4 genes may be related to the effect of calcitonin on the mobility of the PAK osteoclast 4. Zaidi M. et al., Bone 30 (5): 655-63 (2002); Jaffer Z. M. & Chernoff J. Intl. J. Biochem. Cell Biol. 34 (7): 713-7 (2002). These results show the modulating effects of calcitonin on the genes that affect the direct, autocrine, paracrine, and endocrine regulation of osteoblast function (Table 18). These data support the hypothesis that attributes a bone anabolic effect to calcitonin. TABLE 18 Effects on Osteoblasts Coding Gene Function Calcitonin PTS893 from Salmon Cystatin Antagonists B cathepsins; anti-resorption activity Regulation Glycoprotein alpha-2-HS?,?,? autocrine / paracrine of osteoblast function Profiling gene expression of multiple organs in animals treated with salmon cacycitonin. The organs were exposed where changes in expression were seen. B = bone; K = kidney; M = muscle; P = pituitary; L = liver; T = trachea. The results of this example show the modulating effects of calcitonin on the genes that affect the direct, autocrine, paracrine, and endocrine regulation of osteoblast function. These data support the hypothesis that attributes a bone anabolic effect to calcitonin. It is considered that three families of growth factors, transforming growth factor betas (TGF-s), insulin-like growth factors (IGFs), and bone morphogenetic proteins (BMPs), are the main local regulators of osteogenesis. It is thought that bone morphogenetic proteins have their main effects on the replication of early precursor bone cells and the involvement of osteoblasts. In contrast, it is thought that TGB-ps are the most potent inducers of the replication of compromised bone cells and the production of the osteoblast matrix, while IGFs seem to integrate and extend the effect of both factors. McCarthy T. L. et al., Crit. Rev. Oral Biol. Med. 11 (4): 409-22 (2000). These results support the fact that both salmon calcitonin and PTS893 are able to regulate these local and systemic factors involved in bone metabolism. The fact that salmon chloride regulates the a2-HS glycoprotein (AHSG), which blocks TGF-β-dependent signaling in osteotic cells, also supports this role. Mice lacking AHSG display growth plate defects increased bone formation with age, and improved cytokine-dependent osteogenesis. Szweras M. et al., J. Biol. Chem., 27 (22): 19991-19997 (2002). It was also shown that salmon calcitonin and PTS893 modulate the expression of genes encoding vascular endothelial growth factor (VEGF). It is known that VEGF has a key role in normal and pathological angiogenesis. The critical role of angiogenesis for successful osteogenesis during endochondral ossification is well documented. Vascular endothelial growth factor indirectly induces the proliferation and differentiation of osteoblasts by stimulating endothelial cells to produce osteoanabolic growth factors. Wang D. S. et al., Endocrinology 138 (7): 2953-62 (1997). In addition, the vascular endothelial growth factor stimulates the migration of primary human osteoblasts, suggesting a functional role in bone formation and remodeling. ayr-Wohlfahrt U. et al., Bone 30 (3 =: 472-7 (2002).
The effects of parathyroid hormone on osteoblasts to mediate both resorption and bone formation have been widely described. Swarthout J. T. et al., Gene 282 (1-2): 1-17 (2002). Here it was possible to confirm the effect of PTS893 on interleukin 6 (I L-6) cytokines, which mediates the paracrine activation of osteoclast differentiation and activity. Greenfield E. M. et al., Life Sci. 65: 1087-102 (1999). PTS893 also produced a sharp increase over nuclear receptors (steroid / thyroid family). TABLE 19 Genetic Expression Profiling: Growth Factors and Hormones Probe set identifier Times of Controls sCT of expression Gene Coding Mean Average Change GeneChip® 39407 at bone morphogenetic protein 1 448 607 1.36 1122_at chorionic gonadotropin, 263 380 1.44 beta 39945_at polypeptide activation protein of 636 436 -1.46 fibroblasts, alpha 1970_s_at receptor factor 184 108 -1.69 growth of fibroblasts 2 (kinase expressed in bacteria, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome) 32254_at 3 type Follistatin (glycoprotein 1514 2209 1.46 secreted) 38737_at growth factor type 66 37 -1.79 insulin 1 (somatomedin C) 36782_s_at growth factor type 212 323 1.52 insulin 2 (somatomedin A) 1591_s_at growth factor type 293 402 1.37 insulin 2 (somatomedin A ) 40422_at factor binding protein 181 105 -1.73 insulin-like growth 2, 36kDa 37319_at factor binding protein 495 1561 3.15 insulin-like growth 3 1586_at factor binding protein 428 722 1.69 insulin-like growth 3 37319 at protein link factor of 604 879 1.46 growth type insulin 3 1586_at factor binding protein of 355 445 1.25 growth ti po insulin 3 1451_s_at specific factor of 538 292 -1.84 osteoblasts 2 (fasciclin type I), periostin 532_at hormone receptor 1337 1849 1.38 parathyroid 1 234_s_at pleiotrophin (specific factor 710 507 -1.4 of osteoblasts 1) 34820_at pleiotrophin (factor of 422 329 - 1.28 heparin binding growth 8, neurite growth promoting factor 1) 1897_at transcription 1 induced by 176 296 1.68 transforming growth factor beta 1 1385_at growth factor 187 292 1.57 transformant, beta-induced, 68kDa 39588_at superfamily of factor 176 127 -1.39 tumor necrosis (ligand), member 12 31410_at superfamily of factor 197 128 -1.54 tumor necrosis (ligand), member 4 38631_at superfamily of receptor 134 240 1.79 tumor necrosis factor, member 13B 35150_at superfamily of 443 receptor 298 -1.48 tumor necrosis factor, member 5 595_at tumor necrosis factor, 118 191 1.62 protein 3 alpha-induced 1953_at endothelial growth factor 351 557 1.59 vascular 36100_at endothelial growth factor 282 407 1.45 vascular 1953_at endothelial growth factor 521 629 1.21 vascular 37268_at endothelial growth factor 379 504 1.33 vascular B 39091_at response to vitamin A; 421 299 -1.41 related to cytoskeleton Both the caicitonin and parathyroid hormone receptors belong to the superfamily of G-protein receptors. After receptor stimulation, signal transduction is mediated by adenylate cyclase / cAMP / protein kinase, phospholipase phospholipase D, and MAPK (as a late effector) pathways in the case of calcitonin, and by cyclase from adenylate and phospholipase C in the case of the parathyroid hormone. The analysis of genetic profiling allowed us to reconstruct these pathways, showing genes that were modulated by the treatment and that are located at different levels of the signal transduction pathway.
TABLE 20 Effects on Signal Transduction and Cycle C and I u lar Function Coding Gene Calcitonin PTS893 Salmon Transduction of Cyclases of adenylate B signal Calcitin B binding protein Calreticulin B, K, M CREM B, L, PB CDC B kinase, MAPK ALL M MAPK ALL protein kinases Path of phosphatidyl-inositol ALL B Phosphodiesterase (IB, 4A, 4B) ALL B Phospholipase (C, D) ALL B PCNA B Gene expression profiling of multiple organs in animals treated with salmon calcitonin. Organs are exhibited where changes in expression were seen. B = bone; K = kidney; M = muscle; P = pituitary; L = liver; T = trachea.
Salmon calcitonin also seems to exert a direct influence on the cell cycle, since changes in cyclins and cyclin-related proteins could also be observed, as shown in Table 21.
TABLE 21 Profiling Genetic Expression: Signal Transduction The bone morphogenetic protein (BMP) controls the proliferation and differentiation of osteoblasts through Smad proteins. Tob, a member of the emerging family of antiproliferative proteins, is a negative regulator of BMP / Smad signaling in osteoblasts. The Smad pathways were also identified, as well as Tob as one of its regulators, as the genes modulated by the treatment with sCT and PTS893, according to the hypothesized effect of both compounds on the regulation of bone morphogenetic protein on bone remodeling. . Within this context, both compounds seem to exert a direct influence on the cell cycle, since changes in cyclins and cyclin-related proteins could also be observed. Both compounds also regulate the synthesis and degradation of the extracellular matrix components (Table 22), TABLE 22 Effects on the Extracellular Matrix Function Coding Gene Calcitonin PTS893 Salmon Cell binding Integrin B, M, P B Collagenase B signal transduction. Collagen digestion Matrix metalloproteinases 1, II B, L, P, T Synthesis of Endopeptidase / B proteinase collagen procollagen Lysyl hydroxylase B Profiling gene expression of multiple organs in animals treated with salmon calcitonin. Organs are exhibited where changes in expression were seen. B = bone; K = kidney; = muscle; P = pituitary; L = liver; T = trachea.
Salmon calcitonin also regulates the synthesis and degradation of extracellular matrix components, as shown in Table 23.
TABLE 23 Genetic Expression Profiling: Extracellular Matrix Probe set identifier of sCT Controls Expression times Gene Coding Average Average change GeneChip® 36253_at gamma-protein 26305 33265 1.26 bone carboxyglutamate (gla) (osteocalcin) 32094_at sulfotransferase 253 130 -1.95 carbohydrate (chondroitin 6) 3 32094_at sulfotransferase 292 241 -1.21 carbohydrate (chondroitin 6) 3 41447_at carbohydrate synthase 192 107 -1.79 (chondroitin) 1 34042 at condroadherina 7965 10266 1.29 32306_g at collagen, type I, alpha 2 7740 9337 1.21 32488_at collagen, type III, alpha 1 2399 1294 -1.85 (Ehlers-Danlos syndrome type IV, autosomal dominant) 34802 at collagen, type VI, alpha 2 2374 1500 -1.58 bone 1, activation of early T-lymphocytes 1) 38308 at neurocondrina 679 490 -1.39 Of particular interest is the regulation of the Y-frame binding protein (YB-1), which seems to be modulated by both treatments, and in 4 of 6 organs analyzed in the salmon calcitonin group. YB-1 is a protein that interacts with the TGF-β response element in the dista distant region! of the collagen alpha 1 (1) gene. The YB-1 protein activates the collagen promoter, and is translocated in the nucleus during the addition of TGF-β to the fibroblasts, suggesting a role for this protein in TGF-β signaling. Sun W. et al., Matrix Biol. 20 (8); 527-41 (2001).
In addition, salmon calcitonin and PTS893 regulated some aspects of the mineralization of the extracellular bone matrix, because changes were observed in amelogenin, dentine, and in the pituitary ribonucleotide.
TABLE 20 Effects on Ineralization and Visualization Profiling of multiple gene expression in animals treated with salmon calcitonin. The organs where the changes in the expression were seen are exhibited. B = bone; K = kidney; = muscle; P = pituitary; L = liver, T = trachea. All references cited herein are hereby incorporated by reference in their entirety and for all purposes to the same extent as if each publication or patent or individual patent application was indicated in a specific or individual manner as incorporated by reference in its totality for all purposes. In addition, all GenBank accession numbers, Unigene Cluster numbers, and protein accession numbers quoted herein, are hereby incorporated by reference in their entirety and for all purposes to the same degree as if each of these Numbers were indicated in a specific and individual manner as incorporated by reference in their entirety for all purposes. The present invention should not be limited in terms of the particular embodiments described in this application, which are intended as simple illustrations of the individual aspects of the invention. Many modifications and variations can be made without departing from their spirit and scope, as will be apparent to experts in the field. From the foregoing description and accompanying drawings, methods and functionally equivalent apparatuses within the scope of the invention, in addition to those enumerated herein, will be apparent to those skilled in the art. It is intended that these modifications and variations fall within the scope of the appended claims, the present invention should be limited only by the terms of the appended claims, together with the full scope of the equivalents to which these claims are entitled.

Claims (49)

  1. CLAIMS 1. The use of calcitonin in the manufacture of a medicament for the treatment of a condition for which the treatment with an anabolic agent is indicated. 2. The use of claim 1, wherein the condition is atherosclerosis. 3. The use of claim 1 or 2, wherein the calcitonin is salmon calcitonin, 4. The use of calcitonin in the manufacture of a medicament for the treatment of calcium metabolism disorders in a selected patient population, wherein the patient population is selected based on the gene expression profile that indicates the efficacy of calcitonin by the patient to whom the calcitonin is administered. 5. The use of claim 4, wherein the calcitonin is salmon calcitonin. The use of claim 4 or 5, wherein the calcitonin is administered in a therapeutic dose before determining the gene expression profile by the patient. The use of claim 4 or 5, wherein the calcitonin is administered in a s-b-therapeutic dose before determining the gene expression profile by the patient. 8. The use of parathyroid hormone or a parathyroid hormone analogue in the manufacture of a medicament for the treatment of calcium metabolism disorders in a selected patient population, wherein the patient population is selected based on the profile of genetic expression that indicates the effectiveness of the treatment with parathyroid hormone or with the parathyroid hormone analogue by the patient to whom the parathyroid hormone or the parathyroid hormone analogue is administered. 9. The use of claim 8, wherein the parathyroid hormone analogue is PTS893. The use of claim 8 or 9, wherein the parathyroid hormone or the parathyroid hormone analogue is administered in a therapeutic dose before determining the gene expression profile by the patient. The use of claim 8 or 9, wherein the parathyroid hormone or the parathyroid hormone analogue is administered in a subtherapeutic dose before determining the gene expression profile by the patient. 12. A method for treating a condition in a subject, wherein the condition is one for which the administration of a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination thereof is indicated, which comprises the steps of: (a) administering a compound to the subject; (b) obtaining the genetic expression profile of the subject, wherein the gene expression profile comprises the gene expression pattern of one or more genes, wherein the expression patterns of the one or more genes are a consequence of the administration of the compound; and (c) comparing the gene expression profile of the subject to which the compound was administered, with the gene expression profile of a biomarker, indicating the efficacy of the treatment by a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination thereof, wherein a similarity in the gene expression profile of the subject to which the compound was administered, with the gene expression profile of the biomarker, indicates the efficacy of the treatment with the composition. The method of claim 12, wherein the condition is one for which salmon calcitonin is indicated. 14. The method of claim 12, wherein the condition is one for which the PTS893 is indicated. 15. The method of any of claims 12 to 14, wherein the compound administered is a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination thereof. 16. The method of claim 15, wherein the calcitonin is salmon calcitonin. 17. The method of claim 15, wherein the parathyroid hormone analogue is PTS893. 18. The method of any of claims 12 to 17, wherein the subject is a mammal. 19. The method of claim 18, wherein the mammal is a primate. 20. The method of claim 19, wherein the primate is a cinomoigo monkey or a human being. The method of any of claims 12 to 20, wherein the gene expression profile of the biomarker is the gene expression profile of the baseline of the subject prior to administration of the compound. 22. The method of any of claims 20 to 20, wherein the gene expression profile of the biomarker is the gene expression profile or the average gene expression profiles of a vertebrate to which a calcitonin, a hormone, has been administered. parathyroid, a parathyroid hormone analog, or ", a combination thereof. 23. The method of any of claims 12 to 22, wherein the gene expression profile comprises one or more genes selected from the group consisting of isoform a of acid phosphatase 1; 1 receptor type of activin A, type II; Activin A receptor type IIB precursor; activin beta-C chain; alpha 2 HS glycoprotein; amelogenin; annexin V; arylsulfatase precursor E; ATPase H (+) vacuolar; ATPase H (+), vacuolar subunit; ATPase, H + transport, lysosomal; ATPase, transport of H +, lysosomal; ATPase, transport of H +, lysosomal; biglycan; protein m or rf or g e n e t i ca bone 1; protein m or rf or g e n e t i ca bone 10; protein m or rf or g e n eth i ca bone 2A; bone morphogenetic protein 5; precursor of bone morphogenetic protein 6; Calcium binding protein 1 (calbrain); calcium-dependent protein / calmodulin kinase (CaM kinase) II gamma; calreticulin; element modulator that responds to cAMP (CREM); carbonic anhydrase I; carbonic anhydrase II; precursor of oligomeric cartilage matrix protein; cathepsin K; cathepsin W; kinase 1 type CDC; kinase 2 type CDC, isoform hclk2 / 139; chondroitin sulfate proteoglycan 2 (versican); proteoglycan 3 of chondroitin sulfate (neurocan); chorionic somatomammotropin hormone 1; Chymotrypsin C (Caldecrine); type 1 collagen and fusion transcription of PDGFB; collagen type II alpha 1; collagen type III alpha 1; collagen type IV alpha 2; type IX alpha 1 collagen; collagen type VI alpha 1; collagen type VI alpha 2 (AA 570 998); collagen type XI alpha 1; collagen type XI alpha 2; collagen type XI alpha 2; collagen, type I, alpha 2; collagen type IV, alpha 1; collagen type IX, alpha 2; collagen type V, alpha 2; collagen type VI, alpha 1; precursor of collagen type VI, alpha 1; collagen type XVI, alpha 1; collagen type XVI, alpha 1; collagenase 3 (matrix metalloproteinase 13); connective tissue growth factor; cyclin A2; Cyclin B1; Cyclin D2; cyclin E2; cyclin-dependent kinase 5; cyclin-dependent kinase 5, regulatory subunit 1 (p35); cyclin-dependent kinase 6; cyclin 1A-dependent kinase inhibitor (p 21, Cip1); Cystatin B (Stefin B); cytokine-inducible kinase; protein kinase 1 associated with death; protein kinase 3 associated with death; dentin or acidic dentin matrix 1 (DMP1); double specificity phosphatase 9; myotonic protein kinase of dystrophy; pyrophosphatase / ecto-nucleotide phosphodiesterase 1; pyrophosphatase / ecto-nucleotide phosphodiesterase 1; endothelial differentiation, precursor of receptor coupled with G-6 protein, estrogen receptor; estrogen receptor; protein related to estrogen receptor; B-box protein that responds to estrogen (EBBP); fibroblast activation protein; fibroblast growth factor 1 (acid); fibroblast growth factor 18; fibroblast growth factor 4; fibroblast growth factor receptor; Folistatin type 1; Folistatin type 1; glutamate receptor, metabotropic 1; acetyl-glucosaminyl-transferase component GPM N, g i; granulocyte macrophage colony stimulating factor (CSF1); inducible by growth arrest and DNA damage, alpha; protein 10 linked to growth factor receptor; proteoglycan of heparan sulfate 2 (periecan); inositol 1, 4, 5-triphosphate receptor, type 1; 1, 4, 5-triphosphate of inositol receptor, type 1; 1, 4, 5-inositol triphosphate receptor, type 2; kinase 3 isoenzyme of inositol 1,4,5,5-triphosphate; inositol 4-type I beta phosphate phosphatase; inositol 5-phosphate phosphatase; inositol monophosphatase 1 (myo) 1 (or 4), 1; inositol monophosphatase 2 (my) 1 (or 4); insulin-like growth factor (IGF II); insulin-like growth factor 2 (somatomedin A); Insulin-like growth factor binding link; protein of lace 2 of insulin-like growth factor; protein of insulin-like growth factor 3; protein of insulin-like growth factor 5; insulin-like growth factor-binding protein 2; precursor of insulin-like growth factor I I; insulin-like growth factor II precursor; integrin aifa-10 subunit; kinase associated with interleukin 1 receptor; Janus 3 kinase; LIM protein (similar to the binding enigma of rat protein kinase C); Lysyl oxidase type protein; MAD, homologue 3 of mothers against decapentaplégico; AGUKs (membrane-associated guanilate kinase homologs, MAP kinase kinase kinase (TK1), MAPK13, mitogen-activated protein kinase 13, MAPK8IP1: protein-1 interaction with mitogen-activated protein 8 kinase, MEK kinase, metalloproteinase Mitogen-activated protein kinase 1, mitogen-activated protein kinase 8, mitogen-activated protein kinase kinase 1, mitogen-activated protein kinase kinase kinase 4 kinase protein kinase-activated protein kinase 4 activated by mitogen, protein kinase 3 activated by mitogen-activated protein kinase, MDD: associated with monocyte-to-macrophage differentiation, neurocondrine, active T-cell nuclear factor, cytoplasmic, caicyneurin-1-dependent, protein OS 4 (OS 4), osteoblast specific factor 2 OSF 2 (periostin), osteoclast stimulating factor (OSF), PAK4, protein associated with PDFG; kinase of f or sf ati d i I n t osito I 4, catalytic, beta polypeptide; phosphatidylinositol-glycan, class L; Phosphatidylinositol polyphosphate phosphatase 5, isoform B; Phosphatidylinositol 4 phosphate kinase 5, C (1) isoform; Phosphatidylinositol 4 phosphate kinase 5, type I, beta; Phosphatidylinositol 4 phosphate kinase 5, type I I, beta; phosphatidylinositol-glycan, class C (PIG C); f osphodiesterase 4A, specific for cA P; 4D phosphodiesterase, specific for cAMP (drosophil homologous fesf od iesterase (Drosophila)); Phosphodiesterase IB, calmodulin-dependent; Phosphoinositide 3 kinase phosphoinositide 3, catalytic, phosphoinositide 3, class 3 gamma kinase polypeptide; phospholipase C b3 phospholipase C, beta 4; phospholipase D; phosphatidylinositol transfer protein; protein D2 kinase PKD2; p r e p ro-col e g e n o type I, alpha 2; p rep ort I a g e n o type I, alpha 1; pro-collagen alpha 1, type II; Procollagen Lysine Dioxygenase 5; procollagen-proline, 2-oxog lutarate-4-dioxygenase (proline hydroxylase 4), alpha I polypeptide; endometrial protein associated with progestagen (placental protein 14, endometrial alpha 2 globulin associated with pregnancy, alpha uterine protein); prolida (imidodipeptidase) PEPD; proliferating cell nuclear antigen; proylloxy hydroxylase 4 beta; protease, serine, 11 (IGF binding); Proteasome subunit (prosoma, macropain), beta type, 10; activated STAT X protein inhibitor; protein kinase 1 PCTAIRE; substrate 80 H protein kinase C; protein kinase C, alpha; protein kinase, cAMP dependent, catalytic, gamma; protein kinase, cAMP-dependent, regulatory, type I, beta; protein kinase, cAMP-dependent, regulatory, type II, alpha; P2Y purinergic receptor, coupled with G protein, 11; substrate 2 of botulinum toxin C3 related to RAC2 Ras "(rho family, small GTP binding protein Rac2), receptor tyrosine kinase DDR, retinoid receptor X, gamma, ribosomal protein kinase S6, ribosomal protein kinase S6, 9kD , polypeptide 3; SCA P1: secretory carrier membrane protein 1 (vesicular transport); secreted fibrotein 1 (os teo po nt ina, bone sialoprotein I, early T-lymphocyte activation 1), serine proteinase inhibitor (or cysteine) ), cluster H (heat shock protein 47), member 2, serine / threonine kinase 38, protein serine / threonine kinase, SF 1, steroidogenic factor 1, signal transducer and transcription activator 1, signal transducer and transcriptional activator 2, 113kD; signal transducer and transcription activator 5A; signal transducer and transcription activator 5A; signal transducer and transcription activator 6 (STAT6); Smad 3; Smad anchor for activation of the receptor, isoform 1; Smad5; S AD6 (inhibits B P / Smad1 (MADH1)); kinase related to SNF1; SpiB transcription factor (related to SPM / PU.1); Stat5b (stat5b); Serine / threonine kinase related to Ste20; TEIG; early growth response inducible by TGFB; early growth response inducible by TGFB; TIEG; anti-apoptotic factor 1 induced by TGFB1; 12 apoptosis protein induced by TGF beta; TGF beta precursor; TGF beta superfamily protein; Tob; Demellated type 1 kinase; transforming growth factor receptor, beta III (betaglycan, 300kD); Transforming growth factor beta 3 (TGF beta 3); TRIO: triple functional domain (interaction with PTPRF); tubulin alfa 1; tubulin alfa 3; tubulin alfa, H2 alpha isotype; tubulin beta 2; tubulin beta 3; tubulin beta 4; tubulin beta, cofactor D; alpha-2 chain precursor of collagen ti or I; ubiquitin carrier protein E2 C; Vascular endothelial growth factor; Vascular endothelial growth factor; Vascular endothelial growth factor B; and Y-frame link protein 1. The method of claim 23, wherein the gene expression profile comprises an increase in one or more genes selected from the group consisting of bone morphogenetic protein 5; Oligomeric cartilage matrix protein; cathepsin K; Type I collagen pre-pro-alpha-2; and Y-frame binding protein (bone and kidney). 25. The rei indication method 23, wherein the gene expression profile in bone comprises a reduction in one or more genes selected from the group consisting of carbonic anhydrase II; Spi-B; and Y-frame binding protein (muscle). 26. The method of claim 23, wherein the gene expression profile in bone comprises one or more genes selected from the group consisting of PU.1 (SPI1; Spi-B); granulocyte-macrophage colony stimulating factor (CSF1), and associated monocyte-to-macrophage differentiation (M D). The method of claim 23, wherein the gene expression profile in bone comprises a change in the expression of osteoclast stimulating factor (OSF). The method of claim 23, wherein the gene expression profile in bone comprises a change in the expression of vascular endothelial growth factor (VEGF). The method of claim 23, wherein the gene expression profile in bone comprises a change in the expression of a gene selected from the group consisting of integrins; collagenase; matrix metalloproteinases I and II; endopeptidase / procollagen proteinase; Lysyl hydroxylase; aggrecan; precursor of oligomeric cartilage matrix protein; collagens type I, type II, type III, type IV, type V, type VI, type IX, type X, type XI, type XIII, type XIV, type XV, and type XVI); chondroitin sulfate proteoglycan; dermatopontine; proteoglycan of heparan sulfate; and syndecan. The method of claim 23, wherein the gene expression profile in bone comprises a change in the expression of a gene selected from the group consisting of amelogenin; dentine; Ecto-nucleotide pyrophosphatases; and vascular endothelial growth factor. 31. A method for selecting subjects to be included in a clinical study for the determination of the efficacy of a compound, in order to determine the effectiveness of the treatment of a condition, wherein the condition is one for which the administration of a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination thereof, which comprises the steps of: (a) administering a compound to the subject; (b) obtaining the genetic expression profile of the subject, wherein the gene expression profile comprises the gene expression pattern of one or more genes, wherein the expression patterns of the one or more genes are a consequence of the administration of the compound; (c) comparing the gene expression profile of the subject to which the compound was administered, with the gene expression profile of a biomarker; and (d) then: (i) include the subject in the clinical study when the gene expression profile of the subject to which the compound was administered is similar to the biomarker gene expression profile indicating the efficacy of the treatment by a calcitonin , a parathyroid hormone, a parathyroid hormone analog, or a combination thereof; or (ii) exclude the subject from the clinical study when the gene expression profile of the subject to which the compound was administered is different from the biomarker's gene expression profile indicating the efficacy of the treatment by a calcitonin, a parathyroid hormone, a parathyroid hormone analog, or a combination thereof. 32. The method of claim 31, wherein the compound is administered to the subject in a subtherapeutic dose. 33. A method for determining whether a compound has a therapeutic efficacy similar to that of calcitonin, which comprises the steps of: (a) administering a compound to the subject; (b) obtaining the gene expression profile of the subject, wherein the gene expression profile comprises the gene expression pattern of one or more genes, wherein the expression patterns of the one or more genes are a consequence of the administration of the compound; (c) comparing the gene expression profile of the subject to which the compound was administered, with the gene expression profile of a biomarker, indicating the efficacy of the treatment by calcitonin; and (d) then: (i) determining that the compound has a therapeutic efficacy similar to that of calcitonin when the gene expression profile of the subject to which the compound was administered is similar to the gene expression profile of a biomarker of a subject that calcitonin was given; or (ii) determining that the compound has a therapeutic efficacy different from that of calcitonin when the gene expression profile of the subject to which the compound was administered is different from the gene expression profile of the biomarker of a subject to whom calcitonin was administered . 34. The method of claim 33, wherein the calcitonin is salmon calcitonin. 35. The method of reification 33 or 34, wherein the subject is a mammal. 36. The method of claim 35, wherein the mammal is a primate. 37. The method of claim 36, wherein the primate is a cynomolgus monkey or a human being. 38. The method of any of claims 33 to 37, wherein the compound is administered to the subject in a sub-therapeutic dose. 39. A method for determining whether a compound has therapeutic efficacy similar to that of a parathyroid hormone analog, which comprises the steps of: (a) administering a compound to the subject; (b) obtaining the genetic expression profile of the subject, wherein the gene expression profile comprises the gene expression pattern of one or more genes, wherein the expression patterns of the one or more genes are a consequence of the administration of the compound; (c) comparing the gene expression profile of the subject to which the compound was administered, with the gene expression profile of a biomarker, indicating the efficacy of the treatment by a parathyroid hormone analogue; and (d) then: (i) determining that the compound has a therapeutic efficacy similar to that of a parathyroid hormone analogue when the gene expression profile of the subject to which the compound was administered is similar to the gene expression profile of the biomarker of a subject to whom a parathyroid hormone analogue was administered; or (ii) determining that the compound has a therapeutic efficacy different from that of a parathyroid hormone analogue when the gene expression profile of the subject to which the compound was administered is different from the gene expression profile of the biomarker of a subject to which it is assigned. administered a parathyroid hormone analog. 40. The method of claim 39, wherein the parathyroid hormone analogue is PTS893. 41. The method of claim 39 or 40, wherein the subject is a mammal. 42. The method of claim 41, wherein the mammal is a primate. 43. The method of claim 42, wherein the primate is a cynomolgus monkey or a human being. 44. The method of claim 39, wherein the compound is administered to the subject in a subtherapeutic dose. 45. A kit of parts to be used in the determination of the effectiveness of the treatment of a condition for which the administration of a caletonin, a parathyroid hormone, or a parathyroid hormone analog is indicated, which comprises: (a) a reagent for detecting a biomarker of the efficacy of the treatment of a condition for which the administration of a calcitonin, a parathyroid hormone, or a parathyroid hormone analogue is indicated; (b) a container for the reagent; and (c) a product written on or inside the container, which describes the use of the biomarker in the determination of the treatment strategy of the condition. 46. The kit of parts of claim 45, wherein the reagent is a genetic chip. 47. The kit of parts of claim 45, wherein the reagent is a hybridization probe. 48. The kit of parts of claim 45, wherein the reagent is a genetic amplification reagent. 49. The kit of any of the above indications 45 to 48, wherein the biomarker comprises one or more genes selected from the group consisting of isoform a of acid phosphatase 1; 1 receptor type of activin A, type II; Activin A receptor type IIB precursor; activin beta-C chain; alpha 2 HS glycoprotein; amelogenin; annexin V; precursor of arisulfatase E; ATPase H (+) vacuolar; ATPase H (+), vacuolar subunit; ATPase, H + transport, lysosomal; ATPase, transport of H +, lysosomal; ATPase, transport of H +, lysosomal; biglycan; bone morphogenetic protein 1; bone morphogenetic protein 10; bone morphogenetic protein 2A; bone morphogenetic protein 5; precursor of bone morphogenetic protein 6; Calcium binding protein 1 (calbrain); calcium-dependent protein / calmodulin kinase (CaM kinase) II gamma; calreticulin; element modulator that responds to cAMP (CREM); carbonic anhydrase I; carbonic anhydrase II; precursor of oligomeric cartilage matrix protein; cathepsin K; cathepsin W; kinase 1 type CDC; kinase 2 type CDC, isoform hclk2 / 139; p roteog I i ca n 2 of chondroitin sulfate (versican); proteoglycan 3 of chondroitin sulfate (neurocan); chorionic somatomammotropin hormone 1; q u i m i otri s i n a C (caldecrina); type 1 collagen and fusion transcription of PDGFB; collagen type II alpha 1; collagen type MI alpha 1; collagen type IV alpha 2; type IX alpha 1 collagen; collagen type VI alpha 1; collagen type VI alpha 2 (AA 570 998); collagen type XI alpha 1; collagen type XI alpha 2; collagen type XI alpha 2; collagen, type I, alpha 2; collagen type IV, alpha 1; collagen type IX, alpha 2; collagen type V, alpha 2; collagen type VI, alpha 1; precursor of collagen type VI, alpha 1; collagen type XVI, alpha 1; collagen type XVI, alpha 1; collagenase 3 (matrix metalloproteinase 13); connective tissue growth factor; cyclin A2; Cyclin B1; Cyclin D2; cyclin E2; cyclin-dependent kinase 5; cyclin-dependent kinase 5, regulatory subunit 1 (p35); cyclin-dependent kinase 6; inhibitor of cyclin 1A-dependent kinase (p21, Cip1); Cystatin B (Stefin B); cytokine-inducible kinase; protein kinase 1 associated with death; protein kinase 3 associated with death; dentin matrix acid phosphoprotein 1 (DMP1); double specificity phosphatase 9; myobox protein kinase of dystrophy; pyrophosphatase / ecto-nucleotide phosphodiesterase 1; pyrophosphatase / ecto-nucleotide phosphodiesterase 1; endothelial differentiation, precursor of receptor coupled with G-6 protein, estrogen receptor; estrogen receptor; protein related to estrogen receptor; B-box protein that responds to estrogen (EBBP); fibroblast activation protein; fibroblast growth factor 1 (acid); fibroblast growth factor 18; fibroblast growth factor 4; fibroblast growth factor receptor; Folistatin type 1; Folistatin type 1; glutamate receptor, metabotropic 1; acetyl-glucosaminyl-transferase component GPI1 N, gp1; granulocyte macrophage colony stimulating factor (CSF1); inducible by growth arrest and DNA damage, alpha; protein 10 linked to growth factor receptor; proteoglycan of heparan sulfate 2 (perlecan); 1, inositol 4,5-triphosphate receptor, type 1; receptor 1, 4, 5-t r i f o sf atos inositol, type 1; inositol 1, 4,5-triphosphate receptor, type 2; kinase 3 isoenzyme of 1, 4, 5-inositol triphosphate; inositol 4 type I beta polyphosphate phosphatase; Inositol 5 polyphosphate phosphatase; m or n of osf at a s 1 of inositol (mine) 1 (or 4), 1; inositol monophosphatase 2 (my) 1 (or 4); insulin-like growth factor (IGF II); insulin-like growth factor 2 (somatomedin A); Insulin-like growth factor binding protein; Insulin-like growth factor-binding protein 2; Insulin-like growth factor-binding protein 3; insulin-like growth factor binding protein 5; insulin-like growth factor-binding protein 2; precursor of insulin-like growth factor II; insulin-like growth factor II precursor; integrin alpha-10 subunit; kinase associated with interleukin 1 receptor; Janus 3 kinase; LIM protein (similar to the binding enigma of rat protein kinase C); Lysyl oxidase type protein; MAD, homologue 3 of mothers against decapentaplégico; MAGUKs (membrane associated guanylate kinase homologs, MAP kinase kinase kinase (MTK1), APK13, mitogen-activated protein kinase 13, MAPK8IP1: protein-1 interaction with mitogen-activated protein 8 kinase, MEK kinase, metalloproteinase Mitogen-activated protein kinase 1, mitogen-activated protein kinase 8, mitogen-activated protein kinase kinase 1; protein kinase kinase kinase 4 kinase activated by mitogen; protein kinase 2 activated by mitogen-activated protein kinase; protein kinase 3 activated by mitogen-activated protein kinase; MDD: associated with monocyte to macrophage differentiation; neurocondrine; nuclear factor of activated T cells, cytoplasmic, dependent on calcineurin 1; OS 4 protein (OS 4); osteoblast-specific factor 2 OSF 2 (periostin); osteoclast stimulating factor (OSF); PAK4; protein associated with PDFG; Phosphatidylinositol 4 kinase, catalytic, beta polypeptide; phosphatidylinositol-glycan, class L; Phosphatidylinositol polyphosphate phosphatase 5, Soforma B; Phosphatidylinositol 4 phosphate kinase 5, C (1) isoform; Phosphatidylinositol 4 phosphate kinase 5, type i, beta; Phosphatidylinositol 4 phosphate kinase 5, type II, beta; phosphatidylinositol-glycan, class C (PIG C); f or sf od i e st rasa 4A, specific to cAMP; f osf od i esterase 4D, specific for cAMP (f osf od iesterase E3 homologue of dunce (Drosophila)); Phosphodiesterase IB, calmodulin-dependent; phosphoinositide 3 kinase; Phosphoinositide 3 kinase, catalytic, gamma polypeptide; phosphoinositide 3, class 3 kinase; phospholipase C b3; phospholipase C, beta 4; phospholipase D; phosphatidylinositol transfer protein; protein D2 kinase PKD2; pre-pro-collagen type I, alpha 2; pre-pro-collagen type I, alpha 1; pro-collagen alpha 1, type II; Procollagen Lysine Dioxygenase 5; procollagen-proline, 2-oxoglutarate-4-dioxygenase (proline hydroxylase 4), alpha I polypeptide; endometrial protein associated with progestogen (placental protein 14, endometrial alpha 2 globulin associated with pregnancy, alpha uterine protein); prolida (im idodipeptidase) PEPD; proliferating cell nuclear antigen; prolixium hydroxylase 4 beta; protease, serine, 11 (IGF binding); Proteasome subunit (prosoma, macropain), beta type, 10; activated STAT X protein inhibitor; protein kinase 1 PCTAIRE; 80K H substrate protein kinase C; protein kinase C, alpha; protein kinase, cAMP dependent, catalytic, gamma; protein kinase, cAMP-dependent, regulatory, type I, beta; protein kinase, cAMP-dependent, regulatory, type II, alpha; P2Y purinergic receptor, coupled with G protein, 11; substrate 2 of botulinum toxin C3 related to RAC2 Ras (rho family, small GTP binding protein Rac2); receptor tyrosine kinase DDR; retino receptor of X, gamma; ribosomal protein kinase S6; ribosomal protein kinase S6, 9kD, polypeptide 3; SCAMP1: secretory carrier membrane protein 1 (vesicular transport); f secreted osphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1); serine (or cysteine) proteinase inhibitor, cluster H (heat shock protein 47), member 2; sperm kinase a / t reo n a n 38; protein kinase s er i n a / t r eo n a n; SF 1; factor is t e ro i d o g e n i c o 1; signal transducer and transcription activator 1; signal transducer and transcription activator 2, 113kD; signal transducer and transcription activator 5A; signal transducer and transcription activator 5A; signal transducer and transcription activator 6 (STAT6); Smad 3; Smad anchor for receptor activation, isoform 1; Smad5; SMAD6 (inhibits BMP / Smad1 (MADH1)); kinase related to SNF1; transcription factor SpiB (related to SPI1 / PU.1); Stat5b (stat5b); Serine / threonine kinase related to Ste20; TEIG; early growth response inducible by TGFB; early growth response inducible by TGFB; TIEG; anti-apoptotic factor 1 induced by TGFB1; 12 apoptosis protein induced by TGF beta; TGF beta precursor; TGF beta superfamily protein; Tob; Demellated type 1 kinase; transforming growth factor receptor, beta III (betaglycan, 300kD); Transforming growth factor beta 3 (TGF beta 3); TRIO: triple functional domain (interaction with PTPRF); tubulin alfa 1; tubulin alfa 3; tubulin alfa, H2 alpha isotype; tubulin beta 2; tubulin beta 3; tubulin beta 4; tubulin beta, cofactor D; alpha-2 chain precursor of type VI collagen; ubiquitin carrier protein E2 C; Vascular endothelial growth factor; Vascular endothelial growth factor; Vascular endothelial growth factor B; and Y-frame link protein 1. STRUCTURE An analysis of multi-organ genetic profiling of the results of an administration to a subject of salmon calcitonin or a parathyroid hormone analogue provides biomarkers of the efficacy of calcitonin treatment and of the efficacy of treatment with parathyroid hormone or with a parathyroid hormone analog. Biomarkers include the expression profiles of the genes for the Y-frame binding protein, BMPs, FGFs, IGFs, VEGF, g Iicopr ote a-2-IIS (AIISG), OSF, nuclear receptors (family of steroids / thyroid), and others. The results obtained support the anabolic effect of salmon calcitonin on bone metabolism.
MXPA06005950A 2003-11-25 2004-11-24 Biomarkers for the efficacy of calcitonin and parathyroid hormone treatment. MXPA06005950A (en)

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